Revision 1

#9910Store at -20C

1 Kit

(3 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb 4511 20 µl 43 kDa Rabbit IgG
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb 4370 20 µl 44, 42 kDa Rabbit IgG
Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb 4668 20 µl 46, 54 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Phospho-MAPK Family Antibody Sampler Kit provides an economical means of evaluating the phosphorylation state of p38, p44/42, and SAPK/JNK mitogen-activated protein kinases. The kit contains enough primary and secondary antibodies to perform two western blot experiments.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

p44/42 MAPK (Erk1/2), SAPK/JNK, and p38 MAPK function in protein kinase cascades that play a critical role in the regulation of cell growth, differentiation, and control of cellular responses to cytokines and stress. p44/42 MAPK is activated by growth and neurotrophic factors. Activation occurs through phosphorylation of threonine and tyrosine residues (Thr202 and Tyr204 in human Erk1) at the sequence T*EY* by a single upstream MAP kinase kinase (MEK). SAPK/JNK and p38 MAPK are activated by inflammatory cytokines and by a wide variety of cellular stresses. Activation of SAPK/JNK occurs via phosphorylation at Thr183 and Tyr185 by the dual specificity enzyme SEK/MKK4. Both MKK3 and SEK phosphorylate p38 MAPK on tyrosine and threonine at the sequence T*GY* to activate p38 MAP kinase (1-5).

  1. Lewis, T. S. et al. (1998) Adv. Cancer Res. 74, 49-139.
  2. Garrington, T.P. and Johnson, G.L. (1999) Curr. Opin. Cell. Biol. 11, 211-218.
  3. Schaeffer, H.J. and Weber, M.J. (1999) Mol. Cell. Biol. 19, 2435-2444.
  4. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem. Sci. 23, 481-485.
  5. Cobb, M.H. (1999) Prog. Biophys. Mol. Biol. 71, 479-500.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #9910

    Phospho-MAPK Family Antibody Sampler Kit

    Phospho-MAPK Family Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from serum-starved HeLa cells treated with TPA (400 nM, 4 hours) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370. The virtual lane view (left) shows the 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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 2 Expand Image
    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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 3 Expand Image
    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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from COS and 293 cells, untreated or UV-treated, using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (upper) or p38 MAPK Antibody #9212 (lower).
    Phospho-MAPK Family Antibody Sampler Kit: Image 5 Expand Image
    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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 6 Expand Image
    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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 7 Expand Image
    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).
    Phospho-MAPK Family Antibody Sampler Kit: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
    Phospho-MAPK Family Antibody Sampler Kit: Image 9 Expand Image
    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).
    Phospho-MAPK Family Antibody Sampler Kit: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
    Phospho-MAPK Family Antibody Sampler Kit: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
    Phospho-MAPK Family Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded 293T cells untreated (left) or UV-treated (right) using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb.
    Phospho-MAPK Family Antibody Sampler Kit: Image 13 Expand Image
    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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded 293T cell pellets, untreated (left) or UV-treated (right), using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb.
    Phospho-MAPK Family Antibody Sampler Kit: Image 15 Expand Image
    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).
    Phospho-MAPK Family Antibody Sampler Kit: Image 16 Expand Image
    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).
    Phospho-MAPK Family Antibody Sampler Kit: Image 17 Expand Image
    Confocal immunofluorescent analysis of COS cells, untreated (left) or anisomycin-treated (right) using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    Phospho-MAPK Family Antibody Sampler Kit: Image 18 Expand Image
    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).
    Phospho-MAPK Family Antibody Sampler Kit: Image 19 Expand Image
    Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with Anisomycin (25µM, 30 min; green) using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) 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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 20 Expand Image
    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.
    Phospho-MAPK Family Antibody Sampler Kit: Image 21 Expand Image
    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.