Name: Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit
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Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 1 — Simple Western™ analysis of lysates (0.1 mg/mL) from Hela cells using mTOR (7C10) Rabbit mAb #2983. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66-440 kDa separation module.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 2 — Simple Western™ analysis of lysates (0.1 mg/mL) from Jurkat cells treated with Calyculin A (100 uM, 30 min) using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb #4060. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 3 — Simple Western™ analysis of lysates (1.0 mg/mL) from HEK 293 cells treated with UV (50 mJ, 30 min recovery) using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb #4668. The virtual lane view (left) shows two target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 4 — Flow cytometric analysis of Jurkat cells using Akt (pan) (C67E7) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 5 — CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either CREB (48H2) Rabbit mAb or Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using SimpleChIP^® ChIP-seq DNA Library Prep Kit for Illumina^® #56795. The figure shows binding across NR4A3 gene.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 6 — Western blot analysis of extracts from 293, A431, COS, C6, and C2C12 cells, using mTOR (7C10) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 7 — Immunoprecipitation of mTOR protein from MCF-7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900, and lane 3 is mTOR (7C10) Rabbit mAb. Western blot analysis was performed using mTOR (7C10) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 8 — Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 9 — Western blot analysis of extracts from COS cells, untreated or treated with either U0126 #9903 (10 µM for 1h) or TPA #4174 (200 nM for 10 m), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb #4370 and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 10 — Western blot analysis of extracts from 293 cells, untreated or UV-treated, NIH/3T3 cells, untreated or UV-treated and C6 cells, untreated or anisomycin-treated, using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 11 — Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from various cell lines, using Akt (pan) (C67E7) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 12 — Western blot analysis of extracts from serum-starved NIH/3T3 cells, untreated or insulin-treated (150 nM, 5 minutes), alone or in combination with λ-phosphatase, using Phospho-mTOR (Ser2448) (D9C2) XP^® Rabbit mAb (upper) or mTOR (7C10) Rabbit mAb #2983.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 13 — After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO^® is added and emits light during enzyme catalyzed decomposition.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 14 — Western Blot analysis of extracts from SK-N-MC, COS, NIH/3T3, C6 and Drosophila S2 cells, using CREB (48H2) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 15 — Western blot analysis of extracts from SK-N-MC cells, untreated or forskolin- and FGF-treated, using Phospho-CREB (Ser133) (87G3) Rabbit mAb (upper) or CREB (48H2) Rabbit mAb #9197 (lower).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 16 — Western blot analysis of extracts from various tissues using CB1 Receptor (D5N5C) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 17 — Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence^® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence^® mTOR siRNA II (+), using mTOR (7C10) Rabbit mAb #2983 and α-Tubulin (11H10) Rabbit mAb #2125. mTOR (7C10) Rabbit mAb confirms silencing of mTOR expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of mTOR siRNA.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 18 — Immunoprecipitation of phospho-Akt (Ser473) from Jurkat extracts treated with Calyculin A #9902 (100nM, 30 min). Lane 1 is 10% input, lane 2 is Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900. Western blot analysis was performed with Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 19 — Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA #4174 as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 20 — Immunohistochemical analysis of paraffin-embedded 293T cells untreated (left) or UV-treated (right) using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 21 — Immunohistochemical analysis of paraffin-embedded human melanoma using Akt (pan) (C67E7) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 22 — Confocal immunofluorescent analysis of HeLa cells, rapamycin-treated (#9904, 10 nM for 2 hours, left), insulin-treated (150 nM for 6 minutes, middle) or insulin- and λ-phosphatase-treated (right), using Phospho-mTOR (Ser2448) (D9C2) XP^® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 23 — Immunoprecipitation of CREB from SK-N-MC extracts. Lane 1 is CREB (48H2) Rabbit mAb, lane 2 is Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900, and lane 3 is 10% input. Western blot analysis was perfomed using CREB (86B10) Mouse mAb #9104. Anti-mouse IgG, HRP-linked Antibody #7076 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 24 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear staining, using Phospho-CREB (Ser133) (87G3) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 25 — Immunoprecipitation of CB1 Receptor from mouse brain extracts. Lane 1 is 10 % input, lane 2 is Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900, and lane 3 is CB1 Receptor (D5N5C) Rabbit mAb. Western blot analysis was performed using CB1 Receptor (D5N5C) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 26 — Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic localization using mTOR (7C10) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 27 — Immunohistochemical analysis of paraffin-embedded MDA-MB-468 xenograft using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb (left) or PTEN (138G6) Rabbit mAb #9559 (right). Note the presence of P-Akt staining in the PTEN deficient MDA-MB-468 cells.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 28 — Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 29 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb in the presence of control peptide (left) or Phospho-SAPK/JNK (Thr183/Tyr185) Blocking Peptide #1215 (right).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 30 — Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Akt (pan) (C67E7) Rabbit mAb in the presence of control peptide (left) or Akt (pan) Blocking Peptide #1085 (right).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 31 — Immunohistochemical analysis of paraffin-embedded human astrocytoma, using CREB (48H2) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 32 — Immunohistochemical analysis of paraffin-embedded mouse lung using Phospho-CREB (Ser133) (87G3) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 33 — Confocal immunofluorescent analysis of mouse hippocampus: dentate gyrus (left), CA1 region (middle), and cerebellum (right), using CB1 Receptor (D5N5C) Rabbit mAb (green). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 34 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using mTOR (7C10) Rabbit mAb in the presence of control peptide (left) or mTOR Blocking Peptide #1072 (right).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 35 — Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain^® Antibody Diluent #8112 (left) to TBST/5% normal goat serum (right) using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb #4060.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 36 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 37 — Immunohistochemical analysis using Akt (pan) (C67E7) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 38 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using CREB (48H2) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 39 — Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-CREB (Ser133) (87G3) Rabbit mAb in the presence of control peptide (left) or Phospho-CREB (Ser133) Blocking Peptide #1090 (right).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 40 — Immunohistochemical analysis of paraffin-embedded mouse brain using mTOR (7C10) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 41 — Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 42 — Immunohistochemical analysis using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb on SignalSlide^™ Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls #8103 (paraffin-embedded NIH/3T3 cells, treated with U0126 #9903 (left) or TPA #4174 (right).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 43 — Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Akt (pan) (C67E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor^® 555 phalloidin (red). Blue pseudocolor = DRAQ5^™ (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 44 — Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, using CREB (48H2) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 45 — Immunohistochemical analysis of paraffin-embedded SK-N-MC cells, untreated (left) or IBMX- and forskolin-treated (right), showing induced nuclear staining, using Phospho-CREB (Ser133) (87G3) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 46 — Confocal immunofluorescent analysis of mouse embryonic fibroblast (MEF) cells using mTOR (7C10) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 47 — Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb on SignalSlide^® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 48 — Confocal immunofluorescent analysis of Drosophila egg chambers, untreated (top) or λ phosphatase-treated (bottom), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb #4370 (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 49 — Immunohistochemical analysis of paraffin-embedded mouse brain, using CREB (48H2) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 50 — Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma, untreated (left) or lambda phosphatase-treated (right), using Phospho-CREB (Ser133) (87G3) Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 51 — Flow cytometric analysis of A549 cells using mTOR (7C10) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 52 — Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 53 — Confocal immunofluorescent analysis of HT1080 cells, starved overnight then treated with U0126 #9903 (10 uM, 2 h; left) or PDBu (Phorbol 12,13-Dibutyrate) #12808 (100 nM, 15 m; right) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb #4370 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 54 — Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb. (Tissue section courtesy of Dr. Sabina Signoretti, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.)

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 55 — Flow cytometric analysis of Jurkat cells, treated with U0126 (10 µM, 2 hrs; green) or treated with TPA #4174 (200 nM, 30 min; blue) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 56 — Confocal immunofluorescent analysis of mouse cerebellum labeled with CREB (48H2) Rabbit mAb (red) and Neurofilament-L (DA2) Mouse mAb #2835 (green). Blue pseudocolor =DRAQ5^® #4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 57 — Confocal immunofluorescent images of rat dentate gyrus, either sham-operated (left) or 15 min ischemia followed by 30 min (center) and 4 h (right) reperfusion, labeled with Phospho-CREB (Ser133) (87G3) Rabbit mAb (red), Neurofilament-L (DA2) Mouse mAb #2835 (blue) and Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (Alexa Fluor^® 488 Conjugate) #4854.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 58 — Immunohistochemical analysis of paraffin-embedded U-87MG xenograft, untreated (left) or lambda phosphatase-treated (right), using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 59 — Confocal immunofluorescent analysis of SK-N-MC cells showing nuclear stain with CREB (48H2) Rabbit mAb (A, red) compared to an isotype control (B). Blue pseudocolor =DRAQ5^® (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 60 — Confocal microscopic images of SK-N-MC cells showing nuclear stain after 25 minute treatment with Forskolin and IBMX using Phospho-CREB (Ser133) (87G3) Rabbit mAb (left, red) compared to untreated cells (right). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 61 — Flow cytometric analysis of Jurkat cells using CREB (48H2) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 62 — Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with TPA #4174 (500nM) and Ionomycin #9995 (1uM,4hrs;green) using Phospho-CREB (Ser133) (87G3) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 63 — Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor^® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5^®#4084 (fluorescent DNA dye).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 64 — Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 2 hr; blue) using Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor^® 488 Conjugate) #4412 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 65 — Immunoprecipitation of Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) from 3T3 cell extracts. Cells were treated with TPA, (200 nM, 15 min). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP^® Isotype Control #3900, and lane 3 is Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb. Western blot was performed using Phosphop44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (D4W3E) mAb #45262 was used as a secondary antibody.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 66 — Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or CREB (48H2) Rabbit mAb (#9197), using SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP^® ChIP-seq DNA Library Prep Kit for Illumina^® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3, a known target gene of both Phospho-CREB and CREB (see additional figure containing ChIP-qPCR data).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 67 — Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells, treated with Forskolin #3828 (30 μM) for 1h and either 10 μl of CREB (48H2) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729, using SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP^® Human NR4A3 Promoter Primers #4829, and SimpleChIP^® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 68 — Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or CREB (48H2) Rabbit mAb (#9197), using SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP^® ChIP-seq DNA Library Prep Kit for Illumina^® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of both Phospho-CREB and CREB (see additional figure containing ChIP-qPCR data).

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 69 — Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP^® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP^® Human NR4A3 Promoter Primers #4829, and SimpleChIP^® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 70 — CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and Phospho-CREB (Ser133) (87G3) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using SimpleChIP^® ChIP-seq DNA Library Prep Kit for Illumina^® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3 gene.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 71 — CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and Phospho-CREB (Ser133) (87G3) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using SimpleChIP^® ChIP-seq DNA Library Prep Kit for Illumina^® (ChIP-Seq, CUT&RUN) #56795. The figures show binding across chromosome 9 (upper), including NR4A3 (lower) gene.

Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit: Image 72 — CUT&RUN was performed with 293 cells treated with Forskolin #3828 (30 μM) for 1h and either Phospho-CREB (Ser133) (87G3) Rabbit mAb or Rabbit (DA1E) mAb IgG XP^® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers and SimpleChIP^® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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To Purchase # 93521

製品#	サイズ	数量	価格	在庫
93521T	1 Kit (9 x 20 microliters)

Product Includes
Related Products

Product Includes	Quantity	Applications	Reactivity	MW(kDa)	Isotype
CB1 Receptor (D5N5C) Rabbit mAb 93815	20 µl	WB IP IF	H M R	60	Rabbit IgG
Phospho-CREB (Ser133) (87G3) Rabbit mAb 9198	20 µl	WB IHC IF F ChIP C&R	H M R	43	Rabbit IgG
CREB (48H2) Rabbit mAb 9197	20 µl	WB IP IHC IF F ChIP C&R	H M R Mk Dm	43	Rabbit IgG
Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb 4060	20 µl	WB IP IHC IF F	H M R Hm Mk Dm Z B	60	Rabbit IgG
Akt (pan) (C67E7) Rabbit mAb 4691	20 µl	WB IP IHC IF F	H M R Mk Dm	60	Rabbit IgG
Phospho-mTOR (Ser2448) (D9C2) XP^® Rabbit mAb 5536	20 µl	WB IP IF	H M R Mk	289	Rabbit IgG
mTOR (7C10) Rabbit mAb 2983	20 µl	WB IP IHC IF F	H M R Mk	289	Rabbit IgG
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb 4370	20 µl	WB IP IHC IF F	H M R Hm Mk Mi Dm Z B Dg Pg Sc	44, 42	Rabbit IgG
Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb 4668	20 µl	WB IP IHC	H M R Dm Sc	46, 54	Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074	100 µl	WB	Rab		Goat

Kit Usage Information

Protocols

2983: Western Blotting, Immunoprecipitation (Magnetic), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
4060: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
4370: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence*, Flow
4668: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin)
4691: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
5536: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence
7074: Western Blotting
9197: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Immunofluorescence, Flow, ChIP Magnetic, CUT&RUN Assay
9198: Western Blotting, Immunohistochemistry (Paraffin), Immunofluorescence, Immunofluorescence, Flow, ChIP Magnetic, Chromatin IP-seq, CUT&RUN Assay
93815: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence

Product Description

The Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit provides an economical means of detecting the activation of downstream cannabinoid receptor signaling pathways using phospho-specific and control antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb detects endogenous levels of p46 and p54 SAPK/JNK when phosphorylated at Thr183 and Tyr185. It will also react with SAPK/JNK singly phosphorylated at Tyr185. Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb may cross-react with phosphorylated p44/42 or p38 MAP kinases. Phospho-CREB (Ser133) (87G3) Rabbit mAb detects endogenous levels of CREB only when phosphorylated at serine 133. Phospho-CREB (Ser133) (87G3) Rabbit mAb also detects the phosphorylated form of the CREB-related protein, ATF-1. Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Ser473. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb detects endogenous levels of p44 and p42 MAP Kinase (Erk1 and Erk2) when dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), and singly phosphorylated at Thr202. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP^® Rabbit mAb does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinases. Phospho-mTOR (Ser2448) (D9C2) XP^® Rabbit mAb detects endogenous levels of mTOR protein only when phosphorylated at Ser2448.

Source / Purification

CB1 Receptor (D5N5C) Rabbit mAb is produced by immunizing animals with recombinant protein specific to the extracellular domains of mouse CB1 Receptor protein. CREB (48H2) Rabbit mAb is produced by immunizing animals with recombinant protein specific to the amino terminus of human CREB-1 protein. Akt (pan) (C67E7) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy-terminal sequence of mouse Akt. mTOR (7C10) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser2481 of human mTOR. Phosphorylation-specific monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to Thr183/Tyr185 of human SAPK/JNK, Ser133 of human CREB, Ser473 of human Akt, Thr202/Tyr204 of human p44 MAP kinase, and Ser2448 of human mTOR protein.

Background

Cannabinoid receptors mediate a number of physiological processes in the brain ranging from appetite regulation, pain, learning, and memory (1). The major cannabinoid receptors in the brain include CB1 and CB2 receptors, which are G-protein coupled receptors (GPCRs). CB1 interacts with other GPCRs including metabotropic glutamate receptor 1, mGluR1 (2). Endogenous ligands, endocannabinoids, but also exogenously introduced compounds such as tetrahydrocannabinol (THC), activate cannabinoid receptors by promoting the exchange of GDP for GTP, leading to a cascade of signaling pathways that are activated to drive various functions. Some of these functions include neurite outgrowth, inflammation, and transcriptional control (3). Components of this kit are readouts for several downstream signaling components of CB1 receptor and they can also be used as a readout for CB1 activation and function. Cannabinoid receptor function is not limited to brain function but may modulate peripheral functions, including immune responses (4,5).

Pathways

Explore pathways related to this product.

Database Links

UniProt ID: P45983 , P27361 , P42345 , P21554 , P16220 , P31751 , Q9Y243 , P31749

Entrez-Gene Id: 5599 , 5595 , 2475 , 1268 , 1385 , 208 , 10000 , 207

View in PhosphoSitePlus^®

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For Research Use Only. Not for Use in Diagnostic Procedures.

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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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Cannabinoid Receptor 1 Downstream Signaling Antibody Sampler Kit #93521