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9915
Apoptosis Antibody Sampler Kit
Primary Antibodies

Apoptosis Antibody Sampler Kit #9915

IF-IC Image 1

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).

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Western Blotting Image 2

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.

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Western Blotting Image 3

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Caspase-3 siRNA II (+), using Caspase-3 (8G10) Rabbit mAb and α-Tubulin (11H10) Rabbit mAb #2125. Caspase-3 (8G10) Rabbit mAb confirms silencing of caspase-3 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of caspase-3 siRNA.

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Western Blotting Image 4

Western blot analysis of extracts from NIH/3T3 cells, untreated or staurosporine-treated (1 µM), and Jurkat cells, untreated or etoposide-treated (25 µM), using PARP Antibody.

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Western Blotting Image 5

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).

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Flow Cytometry Image 6

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.

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Western Blotting Image 7

Western blot analysis of extracts from Jurkat cells (human), L929 cells (mouse), and C6 cells (rat), untreated or treated with staurosporine or cytochrome c as indicated, using Caspase 9 (C9) Mouse mAb.

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Western Blotting Image 8

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).

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Western Blotting Image 9

Western blot analysis of extracts from Jurkat and A20 cells, untreated (-) or treated with Etoposide #2200 (25 μM, overnight; +), using Caspase-7 (D2Q3L) Rabbit mAb.

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Western Blotting Image 10

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).

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Western Blotting Image 11

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.

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IP Image 12

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.

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Western Blotting Image 13

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Caspase-3 siRNA I (+), using Caspase-3 (8G10) Rabbit mAb and p42 MAPK Antibody #9108. Caspase-3 (8G10) Rabbit mAb confirms silencing of caspase-3 expression, while the p42 MAPK Antibody is used to control for loading and specificity of caspase-3 siRNA.

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IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human tonsil using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.

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IF-IC Image 15

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).

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Flow Cytometry Image 16

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb.

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IHC-P (paraffin) Image 17

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).

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Western Blotting Image 18

Western blot analysis of HeLa (human) and NIH/3T3 (mouse) cell extracts, untreated and treated with 1 μM staurosporine (3 hr) in vivo, using Caspase-3 (8G10) Rabbit mAb.

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IF-IC Image 19

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).

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IHC-P (paraffin) Image 20

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)).

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IP Image 21

Immunoprecipitation of cleaved caspase-3 from Jurkat cell extracts untreated (control) or treated with etoposide (25uM 5hrs) (apoptotic) using Caspase-3 (8G10) Rabbit mAb, and western probed with the same antibody.

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IF-IC Image 22

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).

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IHC-P (paraffin) Image 23

Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

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IHC-F (frozen) Image 24

Immunohistochemical analysis of frozen H1650 xenograft, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

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Flow Cytometry Image 25

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).

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IF-IC Image 26

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).

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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
Caspase-3 (8G10) Rabbit mAb 9665 20 µl
  • WB
  • IP
H M R Mk 17, 19, 35 Rabbit IgG
PARP Antibody 9542 20 µl
  • WB
H M R Mk 89, 116 Rabbit 
Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb 5625 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 89 Rabbit IgG
Caspase-9 (C9) Mouse mAb 9508 20 µl
  • WB
H M R Hm Mk 47/37/35 (H). 49/39/37 (M). 51/40/38 (R). Mouse IgG1
Cleaved Caspase-9 (Asp330) (E5Z7N) Rabbit mAb 52873 20 µl
  • WB
  • IF
H 37 Rabbit IgG
Caspase-7 (D2Q3L) Rabbit mAb 12827 20 µl
  • WB
H M R 20, 35 Rabbit IgG
Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb 8438 20 µl
  • WB
  • IF
  • F
H M R 18 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

The Apoptosis Antibody Sampler Kit provides an economical means to evaluate the levels of inactive and active caspases. The kit contains enough primary and secondary antibodies to perform two Western blot experiments with each antibody.

Each antibody in the Apoptosis Antibody Sampler Kit detects its respective target at endogenous levels. Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb, Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb, Cleaved Caspase-9 (Asp330) (E5Z7N) Rabbit mAb, and Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb detect only the large cleaved fragments of their respective targets. Caspase-3 (8G10) Rabbit mAb, Caspase-7 (D2Q3L) Rabbit mAb, Caspase-9 (C9) Mouse mAb, and PARP Antibody detect both the full length and the large cleaved fragments of their respective targets.

Monoclonal and polyclonal antibodies are produced by immunizing animals with recombinant human caspase-9 protein or with synthetic peptides corresponding to residues surrounding Pro158 of human caspase-7 protein or the proteolytic cleavage sites of human caspase-3, caspase-9, and PARP proteins. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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 the FasL, TNF-α, DNA damage and ER stress. Fas and TNFR activate caspases 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 the IAPs on caspases (6).

  1. Baker, S.J. and Reddy, E.P. (1998) Oncogene 17, 3261-3270.
  2. Budihardjo, I. et al. (1999) Annu. Rev. Cell Dev. Biol. 15, 269-290.
  3. Nakagawa, T. et al. (2000) Nature 403, 98-103.
  4. Deveraux, Q. L. et al. (1998) EMBO J. 17, 2215-2223.
  5. Li, F. et al. (1998) Nature 396, 580-584.
  6. Du, C. et al. (2000) Cell 102, 33-42.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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