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48734
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit #48734

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Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 1
Western blot analysis of extracts from mouse bone marrow derived macrophages (mBMDM), untreated (-) or treated with Lipopolysaccharides (LPS) #14011 (50 ng/ml, 4 hr; +) followed by Nigericin (sodium salt) #66419 (15 μM, indicated times; +), using Cleaved Gasdermin D (Asp276) (E3E3P) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 2
Western blot analysis of extracts from MEFs from wild-type or SQSTM1/p62 knockout mice using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). MEF SQSTM1/p62 KO cells were kindly provided by Dr. Junying Yuan, Harvard Medical School, Boston MA.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 3
Western blot analysis of extracts from C2C12 cells, untreated (-) or treated with Chloroquine #14774 (50 μM, overnight) using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 4
Western blot analysis of extracts from various cell lines using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 5
Western blot analysis of extracts from MEFs, untreated (-) or treated with Earles Basic Salt Solution (EBSS; 4 hr; +) using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 6
Immunoprecipitation of SQSTM1 from L-929 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific). Western blot was performed using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific). Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 7
Immunohistochemical analysis of paraffin-embedded MEF wild-type cell pellet (left, positive) or MEF SQSTM1/p62 KO cell pellet (right, negative) using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific). MEF SQSTM1/p62 KO cells were kindly provided by Dr. Junying Yuan, Harvard Medical School, Boston MA.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 8
Immunohistochemical analysis of paraffin-embedded mouse forestomach using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 9
Immunohistochemical analysis of paraffin-embedded mouse kidney using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 10
Immunohistochemical analysis of paraffin-embedded mouse spleen using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 11
Immunohistochemical analysis of paraffin-embedded rat spleen using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 12
Immunohistochemical analysis of paraffin-embedded mouse small intestine using SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 13
Confocal immunofluorescent analysis of wild-type MEFs, either untreated (left) or treated with Chloroquine #14774 (50 μM, 18 hours; center), and SQSTM1/p62 knock-out MEFs treated with chloroquine (right), using SQSTM1 (D6M5X) Rabbit mAb (green). Actin filaments were labeled with β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). MEF SQSTM1/p62 KO cells were kindly provided by Dr. Junying Yuan, Harvard Medical School, Boston MA.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 14
Western blot analysis of L-929 cells, untreated (-), or treated with combinations of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/ml, 4 hr; +), SM-164 (100 nM, 4 hr; +), and necrostatin-1 (Nec-1, 50 μM, 4 hr; +), using Phospho-MLKL (Ser345) (D6E3G) Rabbit mAb (upper), total MLKL (D6W1K) Rabbit mAb (Mouse Specific) #37705 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 15
Confocal immunofluorescent analysis of L-929 cells, untreated (left), pre-treated with Z-VAD (20 μM, 30 min) followed by treatment with SM-164 (100 nM) and Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/mL, 2.5 hr; center) and then post-processed with λ-phosphatase (right), using Phospho-MLKL (Ser345) (D6E3G) Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087 (fluorescent DNA dye).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 16
Western blot analysis of extracts from wild-type (WT) MEF or MEF/RIPK1 knockout (KO) cells, untreated (-) or treated with Z-VAD(OMe)-FMK #60332 (20 μM, 30 min) followed by Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/ml, 7 hr) and SM-164 (100 nM, 7 hr), using Phospho-RIP (Ser166) (E7G6O) Rabbit mAb (upper), RIP (D94C12) XP® Rabbit mAb #3493 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). MEF/RIPK1 KO cells were kindly provided by Dr. Junying Yuan, Harvard Medical School, Boston, MA.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 17
Western blot analysis of extracts from L-929 cells, untreated (-) or treated with Z-VAD(OMe)-FMK #60332 (20 μM, 30 min) followed by Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/ml, indicated times) and SM-164 (100 nM, indicated times), using Phospho-RIP (Ser166) (E7G6O) Rabbit mAb (upper), RIP (D94C12) XP® Rabbit mAb #3493 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 18
Western blot analysis of extracts from H9c2(2-1) cells, untreated (-) or treated with Z-VAD(OMe)-FMK #60332 (20 μM, 30 min) followed by Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/ml, 4.5 hr) and SM-164 (100 nM, 4.5 hr), using Phospho-RIP (Ser166) (E7G6O) Rabbit mAb (upper), RIP (D94C12) XP® Rabbit mAb #3493 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 19
Immunoprecipitation of phospho-RIP (Ser166) protein from L-929 cell extracts treated with Z-VAD(OMe)-FMK #60332 (20 μM, 30 min) followed by Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/ml, 2 hr) and SM-164 (100 nM, 2 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 Phospho-RIP (Ser166) (E7G6O) Rabbit mAb. Western blot analysis was performed using Phospho-RIP (Ser166) (E7G6O) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 20
Western blot analysis of cell extracts and media from mouse bone marrow derived macrophages (mBMDM), untreated (-), or treated (+) with combinations of LPS #14011 (50 ng/ml, 4 hr) followed by nigericin (15 μM, 45 min) using Cleaved-IL-1β (Asp117) (D7V2A) Rabbit mAb (Mouse Specific) (upper) or total IL-1β (D3H1Z) Rabbit mAb (Mouse Specific) #12507 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 21
Immunoprecipitation of Cleaved-IL-1β (Asp117) from extracts of media from mouse bone marrow derived macrophages (mBMDM) treated with LPS #14011 (50 ng/ml, 4 hr) followed by nigericin (15 μM, 45 min). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cleaved-IL-1β (Asp117) (E7V2A) Rabbit mAb (Mouse Specific). Western blot was performed using Cleaved-IL-1β (Asp117) (E7V2A) Rabbit mAb (Mouse Specific). Anti-Rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 22
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.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 23
Western blot analysis of extracts from HCT 116 and HCT 116 LC3B knockout cells, untreated (-) or treated with Chloroquine #14774 (50 μM, 18 hr) using LC3B (E5Q2K) Mouse mAb #83506 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the HCT 116 knockout cells confirms the specificity of the antibody for LC3B.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 24
Western blot analysis of extracts from HeLa, C2C12, and KNRK cells, untreated (-) or treated with Chloroquine (50 μM, overnight; +) #14774 using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 25
Western blot analysis of extracts from HeLa cells or HeLa cells with a knockout of LC3B (HeLa/LC3B KO) using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 26
Western blot analysis of extracts from A549 cells, untreated (-) or starved with Earle's Balanced Salt Solution (EBSS) (indicated times) using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 27
Western blot analysis of extracts from MCF7 and Raji cells, untreated (-) or treated with Torin 1 (250 nM, 5 hr; +) #14379, using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 28
Immunoprecipitation of LC3B from HeLa cells treated with Chloroquine (50 μM, overnight) # 14774. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control, and lane 3 is LC3B (E5Q2K) Mouse mAb. Western blot was performed using LC3B (E5Q2K) Mouse mAb. Anti-mouse IgG, HRP-linked Antibody #7076 was used as a secondary antibody.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 29
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 30
Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded HCT116 cell pellets, untreated (left-top) or treated with Chloroquine #14774 (right-top), and HCT116 LC3B knockout cell pellets, untreated (left-bottom) or treated with Chloroquine #14774 (right-bottom), using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 34
Immunohistochemical analysis of paraffin-embedded normal human brain using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 35
Immunohistochemical analysis of paraffin-embedded KARPAS 299 cell pellet (left, high-expressing) or VCaP cell pellet (right, low-expressing) using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 36
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or treated with Chloroquine #14774 (right), using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 37
Immunohistochemical analysis of paraffin-embedded normal human spleen using LC3B (E5Q2K) Mouse mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 38
Confocal immunofluorescent analysis of HCT 116 cells either untreated (left) or treated with Chloroquine #14774 (50 µM, overnight) (center) or LC3B HCT 116 knockout cells treated with Chloroquine #14774 (50 µM, overnight) (right) using LC3B (E5Q2K) Mouse mAb (green). Actin filaments were labeled with β-Actin (13E5) Rabbit mAb (red) and nuclei were labeled with DAPI #4083 (blue).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 39
Flow cytometric analysis of HCT-116 cells, wild-type (green, high expression) or LC3B knockdown (blue, negative expression), using LC3B (E5Q2K) Mouse mAb or a concentration-matched Mouse (E7Q5L) mAb IgG2b Isotype Control #53484 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 40
Western blot analysis of L-929 cells, untreated (-), or treated with combinations of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), SM-164 (100 nM, 3 hr; +), and mouse TNF-α (20 ng/ml, 3 hr; +), using Phospho-RIP3 (Thr231/Ser232) (E7S1R) Rabbit mAb (upper), RIP3 (D8J3L) Rabbit mAb #15828 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 41
Confocal immunofluorescent analysis of L-929 cells, untreated (left), pre-treated with Z-VAD (20 μM, 30 min) followed by treatment with SM-164 (100 nM) and Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/mL, 2.25 hr; center), or pre-treated with Z-VAD followed by treatment with SM-164 and hTNF-α and post-processed with λ-phosphatase (right), using Phospho-RIP3 (Thr231/Ser232) (E7S1R) Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 42
Western blot analysis of extracts from serum-starved Neuro-2a cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 43
Western blot analysis of extracts from serum-starved H-4-II-E cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 6 hr; +), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 44
Immunoprecipitation of Cleaved PARP (Asp214) from Neuro-2a cell extracts treated with Staurosporine #9953 (1 μM, 3 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific). Western blot was perform using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 45
Immunohistochemical analysis of paraffin-embedded 3T3 cell pellet, untreated (left, negative) or treated with Staurosporine #9953 (right, positive), using Cleaved PARP (Asp214)(D6X6X) mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 46
Immunohistochemical analysis of paraffin-embedded E14 rat embryo using Cleaved PARP (Asp214)(D6X6X) mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 47
Immunohistochemical analysis of paraffin-embedded H-4-II-E cell pellet, untreated (left, negative) or treated with Staurosporine #9953 (right, positive), using Cleaved PARP (Asp214)(D6X6X) mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 48
Immunohistochemical analysis of paraffin-embedded mouse ovary using Cleaved PARP (Asp214)(D6X6X) mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 49
Immunohistochemical analysis of paraffin-embedded mouse spleen using Cleaved PARP (Asp214)(D6X6X) mAb (Rodent Specific).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 50
Confocal immunofluorescent analysis of Neuro-2a cells, untreated (left, negative) or treated with Staurosporine #9953 (1 μM, 3 hr; right, positive), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 51
Flow cytometric analysis of serum-starved Neuro-2a cells, untreated (blue) or treated with Staurosporine #9953 (1 μM, 3 hr; green), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific) (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.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 52
Flow cytometric analysis of serum-starved H-4-II-E cells, untreated (blue) or treated with Staurosporine #9953 (1 μM, 3 hr; green), using Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific) (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.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 53
Western blot analysis of extracts from C6 (rat), NIH/3T3 (mouse), and Jurkat (human) cells, untreated or treated with staurosporine #9953 (1uM, 3hrs) or etoposide #2200 (25uM, 5hrs) as indicated, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 54
Immunoprecipitation of extracts from Jurkat cells, untreated or etoposide-treated (25uM, 5hrs), using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb. Western blot was performed using the same antibody.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 55
Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb in the presence of control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide (#1050) (right).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 56
Immunohistochemical analysis using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb on SignalSlide® Cleaved Caspase-3 IHC Controls #8104 (paraffin-embedded Jurkat cells, untreated (left) or etoposide-treated (right)).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 57
Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 58
Confocal immunofluorescent images of HT-29 cells, untreated (left) or Staurosporine #9953 treated (right) labeled with Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit: Image 59
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with etoposide #2200 (green), using Cleaved Caspase-3(Asp175) (5A1E) Rabbit mAb compared to a nonspecific negative control antibody (red).
To Purchase # 48734T
製品番号 サイズ 価格 在庫
48734T
1 Kit  (9 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb 9664 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk 17, 19 Rabbit IgG
Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific) 94885 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
M R 89 Rabbit IgG
Phospho-RIP (Ser166) (E7G6O) Rabbit mAb 53286 20 µl
  • WB
  • IP
M R 78 Rabbit IgG
Phospho-RIP3 (Thr231/Ser232) (E7S1R) Rabbit mAb 91702 20 µl
  • WB
  • IF
M 46-62 Rabbit IgG
Phospho-MLKL (Ser345) (D6E3G) Rabbit mAb 37333 20 µl
  • WB
  • IF
M 54 Rabbit IgG
Cleaved Gasdermin D (Asp276) (E3E3P) Rabbit mAb 10137 20 µl
  • WB
M 31 Rabbit IgG
Cleaved-IL-1β (Asp117) (E7V2A) Rabbit mAb (Mouse Specific) 63124 20 µl
  • WB
  • IP
M 17 Rabbit IgG
LC3B (E5Q2K) Mouse mAb 83506 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 14, 16 Mouse IgG2b
SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific) 23214 20 µl
  • WB
  • IP
  • IHC
  • IF
M R 62 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit provides an economical means of detecting common readouts in apoptosis, necroptosis, pyroptosis, and autophagy. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Mouse Reactive Cell Death and Autophagy Antibody Sampler Kit detects endogenous levels of its target protein. Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb detects endogenous levels of the large fragment (17/19 kDa) of activated caspase-3 resulting from cleavage adjacent to Asp175. This antibody does not recognize full-length caspase-3 or other cleaved caspases. Cleaved PARP (Asp214) (D6X6X) Rabbit mAb (Rodent Specific) recognizes endogenous levels of the large fragment (89 kDa) of rodent PARP only when cleaved at Asp214. Cleaved Gasdermin D (Asp276) (E3E3P) Rabbit mAb recognizes endogenous levels of the amino fragment of mouse Gasdermin D protein only when cleaved at Asp276. Cleaved-IL-1β (Asp117) (E7V2A) Rabbit mAb (Mouse Specific) recognizes endogenous levels of mouse IL-1β protein only when cleaved at Asp117. Phospho-RIP3 (Thr231/Ser232) (E7S1R) Rabbit mAb recognizes endogenous levels of RIP3 protein only when phosphorylated at Thr231/Ser232. This antibody may not recognize RIP3 when only singly phosphorylated at Thr231 or Ser232. Phospho-RIP (Ser166) (E7G6O) Rabbit mAb recognizes endogenous levels of RIP protein only when phosphorylated at Ser166. Phospho-MLKL (Ser345) (D6E3G) Rabbit mAb recognizes endogenous levels of mouse MLKL protein only when phosphorylated at Ser345. Weak, non-specific nuclear staining has been observed by immunofluorescence (IF-IC). LC3B (E5Q2K) Mouse mAb detects both type I and type II forms of LC3B. Cross reactivity was not detected with other family members. SQSTM1/p62 (D6M5X) Rabbit mAb (Rodent Specific) recognizes endogenous levels of total rodent SQSTM1/p62 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp175 of human caspase-3, Asp214 of rodent PARP, Asp276 of mouse Gasdermin D, Asp117 of mouse IL-1β, Gly300 of mouse SQSTM1/p62, residues near the amino terminus of human LC3B, and synthetic phosphopeptides corresponding to Ser166 of mouse RIP, Thr231/Ser232 of mouse RIP3, and Ser345 of mouse MLKL.

Background

Regulated cell death has been classified based on distinct morphological and biochemical pathways (1). Type I cell death, or apoptosis, is characterized by cytoplasmic shrinkage, chromatin condensation, nuclear fragmentation, plasma membrane blebbing, and phagocytic update of dead cells. Apoptosis can occur through extrinsic pathways involving extracellular factors, including the activation of death receptors, or through intrinsic pathways involving intracellular perturbations, including mitochondrial outer membrane permeabilization (2). Both of these apoptotic pathways lead to activation of caspases, a family of cysteine acid proteases that are synthesized as inactive zymogens containing pro-domains, followed by large (p20) and small (p10) subunits which are proteolytically activated in a cascade-like fashion. Caspase-3 is a key downstream protease activated by both extrinsic and intrinsic apoptotic pathways and cleaves a large number of proteins involved in the disassembly of the cell, including poly(ADP-ribose) polymerase (PARP), a protein involved in the DNA damage response.   Type II cell death, or autophagy, manifests with extensive cytoplasmic vacuolization, and like apoptosis, can include phagocytic update. Autophagy is a catabolic process for the degradation of cellular components including protein aggregates, damaged organelles, and pathogens (3). The process involves the engulfment of these components into a double membrane structure, the autophagosome, which fuses to the lysosome for degradation. Autophagy requires, and can be monitored by, the conversion of LC3 family members, such as LC3B, from a type I form to a lipidated type II form that is incorporated into the autophagosome membrane and binds to a variety of cargo receptors. Cargo receptors such as SQSTM1/p62 bind LC3 along with ubiquitinated proteins that are targeted for degradation. SQSTM1/p62 is also degraded during this process, and thus its expression is frequently used to monitor this process. Type III cell death, or necrosis, manifests with plasma membrane permeability with cellular swelling and fragmentation, and lacks a clear phagocytic response which then leads to an inflammatory signaling with the release of damage-associated molecular patterns (DAMPs). Necrosis can be triggered by multiple regulated pathways including necroptosis and pyroptosis. Necroptosis is regulated by the kinase activities of RIP and RIP3 and the pore forming ability of MLKL (4). Necroptosis requires the activation of RIP3 which then phosphorylates MLKL at Ser358 (Ser345 in mouse). Phosphorylation of MLKL leads to generation of a pore complex involved in cell swelling and the secretion of DAMPs. RIP3 activation is triggered through several RIP homotypic interaction motif (RHIM) domain interactions including RIP, TRIF, and ZBP1 and results in the phosphorylation of RIP3 at Ser227 (Thr231/Ser232 in mouse). Canonical necroptosis signaling is mediated by RIP, and this can be inhibited by necrostatins, small molecules that directly inhibit RIP kinase activity. Activation of RIP can be monitored through autophosphorylation sites including Ser166. Pyroptosis is generally induced in cells of the innate immune system, and is characterized by cleavage of Gasdermin D (5). The amino-terminal fragment of Gasdermin D produced following cleavage by inflammatory caspases (Caspase-1, -4, -5), oligomerizes to form a pore. Canonical cleavage of Gasdermin D occurs through a two-step process. The first step involves transcriptional regulation of targets such as NLRP3 and the pro-forms of IL-1β and IL-18. In the second execution step, Caspase-1 is activated through formation of inflammasome complexes. Activated Caspase-1 cleaves Gasdermin D as well as IL-1β and IL-18 to their mature forms, and these active cytokines are secreted through pores formed by Gasdermin D.
  1. Galluzzi, L. et al. (2018) Cell Death Differ 25, 486-541.
  2. Green, D.R. (1998) Cell 94, 695-8.
  3. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
  4. Shan, B. et al. (2018) Genes Dev 32, 327-40.
  5. Shi, J. et al. (2017) Trends Biochem Sci 42, 245-54.

Pathways & Proteins

Explore pathways + proteins related to this product.

使用に関する制限

法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

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