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9929
Cleaved Caspase Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Cleaved Caspase Antibody Sampler Kit #9929

Citations (18)
Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells treated with Etoposide (25 μM, 5 hours) using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625. 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.
Simple Western™ analysis of lysates (0.1 mg/mL) from Jurkat cells treated with Cytochrome C using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664. 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.
Western blot analysis of extracts from HeLa and Jurkat cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +) or Etoposide #2200 (25 μM, overnight; +), using Cleaved Caspase-9 (Asp330) (E5Z7N) Rabbit mAb (upper), Caspase-9 (C9) Mouse mAb #9508 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HeLa cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr), Jurkat cells, untreated or etoposide-treated (25 μM, overnight), and THP-1 cells, untreated or cycloheximide-treated (CHX, 10 μg/ml, overnight) followed by treatment with TNF-α #8902 (20 ng/ml, 4 hr), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (upper), or total PARP Antibody #9542 (lower).
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.
Western blot analysis of extracts from HeLa, C2C12, and H-4-II-E cells, untreated (-) or treated (+) with Staurosporine #9953 (1 μM, 3 hr), using Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb (upper) and Caspase-7 Antibody #9492 (lower).
Western blot analysis of extracts from Jurkat cells, untreated or cytochrome c-treated (0.25 mg/ml), and HeLa cells, untreated, staurosporine-treated (1 µM), or cytochrome c-treated (0.25 mg/ml), using Cleaved Caspase-9 (Asp315) Antibody (upper) or Caspase-9 Antibody #9502 (lower).
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.
Western blot analysis of extracts from various cell lines, untreated, cytochrome c (1hr, 0.25 mg/ml in vitro), or raptinal (10 μM, 2 hr) treated using Cleaved Caspase-6 (Asp162) Antibody #9761 (upper), Caspase-6 Antibody #9762 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (1 μM, 3 hr; right), using Cleaved Caspase-9 (Asp330) (E5Z7N) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human tonsil using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (1 μM, 3 hr; right), using Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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.
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (green). Actin filament were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb.
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with Etoposide #2200 (25 uM, 18 hr; green) using Cleaved PARP (Asp214) (D64E10) 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.
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).
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)).
Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.
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).
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 # 9929
Cat. # Size Qty. Price
9929T
1 Kit  (6 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 Caspase-6 (Asp162) Antibody 9761 20 µl
  • WB
H M R 18 Rabbit 
Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb 8438 20 µl
  • WB
  • IF
  • F
H M R 18 Rabbit IgG
Cleaved Caspase-9 (Asp330) (E5Z7N) Rabbit mAb 52873 20 µl
  • WB
  • IF
H 37 Rabbit IgG
Cleaved Caspase-9 (Asp315) Antibody 9505 20 µl
  • WB
  • IP
H 35 Rabbit 
Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb 5625 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 89 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Cleaved Caspase Antibody Sampler Kit provides an economical means to evaluate the activation status of caspases by detecting their cleaved forms. The kit contains enough primary and secondary antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Cleaved Caspase-3 (Asp175), Cleaved Caspase-6 (Asp162), Cleaved Caspase-7 (Asp198), Cleaved Caspase-9 (Asp315), Cleaved Caspase-9 (Asp330), and Cleaved PARP (Asp214) Antibodies detect endogenous levels of the large cleavage fragments of their respective targets. These antibodies will not cross-react with their respective full-length proteins.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino-terminal residues adjacent to Asp175 of human caspase-3 or to residues surrounding Asp214 in human PARP, Asp330 of human caspase-9, or Asp198 of human caspase-7. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp315 of human caspase-9 or to the carboxy-terminal sequence of the large subunit (p18) of rat caspase-6. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Apoptosis is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 8, 9, 10, and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6, and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF, and lamin A and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase (1). Proapoptotic stimuli include FasL, TNF-α, DNA damage and ER stress. Fas and TNFR activate caspase-8 and -10 (2), DNA damage leads to the activation of caspase-9 and ER stress leads to the calcium-mediated activation of caspase-12 (3). The inhibitor of apoptosis protein (IAP) family includes XIAP and survivin and functions by binding and inhibiting several caspases (4,5). Smac/Diablo, a mitochondrial protein, is released into the cytosol upon mitochondrial stress and competes with caspases for binding of IAPs. The interaction of Smac/Diablo with IAPs relieves the inhibitory effects of IAPs on caspases (6).

Limited Uses

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