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9930
Apoptosis Antibody Sampler Kit (Mouse Preferred)

Apoptosis Antibody Sampler Kit (Mouse Preferred) #9930

Western Blotting Image 1

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 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 BaF3, Raw264.7 and C2C12 cell lines, using Caspase-8 Antibody (Mouse Specific).

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Western Blotting Image 4

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct overexpressing mouse caspase-8 (mCasp8) (+), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific).

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Western Blotting Image 5

Western blot analysis of extracts from L929 cells, untreated or Brefeldin A-treated (10 µM, 30 hours), using Caspase-12 Antibody.

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Western Blotting Image 6

Western blot analysis of extracts from NIH/3T3 cells, untreated, staurosporine-treated (1 µM), or cytochrome c-treated (0.25 mg/ml), using Caspase-9 Antibody. p39: caspase-9 cleaved at Asp368. p37: caspase-9 cleaved at Asp353.

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Western Blotting Image 7

Western blot analysis of extracts from L929 cells, untreated or staurosporine-treated, using Cleaved Caspase-9 (Asp353) Antibody (upper) or Caspase-9 Antibody #9504 (lower).

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Western Blotting Image 8

Western blot analysis of NIH/3T3 cells, untreated or staurosporine-treated (1 µM), using Cleaved PARP (Asp214) (7C9) Mouse mAb (Mouse Specific).

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Western Blotting Image 9

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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Western Blotting Image 10

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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IP Image 11

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 12

Western blot analysis of extracts from CTLL-2 cells, untreated or treated with cycloheximide (CHX, 10 μg/ml, overnight) followed by hTNF-α #8902 (20 ng/ml, 4 hours), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific).

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IF-IC Image 13

Confocal immunofluorescent analysis of NIH/3T3 cells, staurosporine-treated (left) or untreated (right), using Cleaved Caspase-9 (Asp353) Antibody (Mouse Specific) (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Western Blotting Image 14

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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IHC-P (paraffin) Image 15

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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Flow Cytometry Image 16

Flow cytometric analysis of CTLL-2 cells, untreated (red) or treated with cyclohexamide and hTNF-α #8902 (blue), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific).

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IP Image 17

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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IHC-P (paraffin) Image 18

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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IF-IC Image 19

Confocal immunofluorescent analysis of Raw 264.7 cells, untreated (left) or treated with TNF-α and Cycloheximide (20 ng/ml and 2.5 ug/ml, 4 hours and 3 hours; right), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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IHC-P (paraffin) Image 20

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 21

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

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Flow Cytometry Image 22

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 23

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
Caspase-3 (8G10) Rabbit mAb 9665 20 µl
  • WB
  • IP
H M R Mk 17, 19, 35 Rabbit IgG
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb 9664 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk 17, 19 Rabbit IgG
Caspase-8 Antibody (Mouse Specific) 4927 20 µl
  • WB
M 45, 57 Rabbit 
Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific) 8592 20 µl
  • WB
  • IP
  • IF
  • F
M 18, 43 Rabbit IgG
Caspase-12 Antibody 2202 20 µl
  • WB
M 42, 55 Rabbit 
Caspase-9 Antibody (Mouse Specific) 9504 20 µl
  • WB
M 37, 39, 49 Rabbit 
Cleaved Caspase-9 (Asp353) Antibody (Mouse Specific) 9509 20 µl
  • WB
  • IF
M 37 Rabbit 
Cleaved PARP (Asp214) (7C9) Mouse mAb (Mouse Specific) 9548 20 µl
  • WB
M 89 Mouse IgG2b
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 (Mouse Specific) is designed for use with mouse samples and offers an economical means to evaluate the levels of active and inactive 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 (Mouse Preferred) recognizes endogenous levels of its respective target. Caspase-3 (8G10) Rabbit mAb detects full-length (35 kDa) and the large fragment (17/19 kDa) of caspase-3 resulting from cleavage at aspartic acid 175. Cleaved Caspase-3 (Asp175) (5A1) Rabbit mAb detects the large fragment (17/19 kDa) of caspase-3 resulting from cleavage adjacent to Asp175. Caspase-8 Antibody (Mouse Specific) detects full length (57 kDa) and the cleaved intermediate (p43) of mouse caspase-8. Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb (Mouse Specific) detects the p18 subunit of mouse caspase-8 resulting from cleavage at Asp387and the cleavage product containing the pro-domain (p43). Caspase-9 Antibody (Mouse Specific) detects both full length (49 kDa) and the large fragments of mouse caspase-9 resulting from cleavage at aspartic acid 353 (37 kDa) and/or aspartic acid 368 (39 kDa). Cleaved Caspase-9 (Asp353) Antibody (Mouse Specific) detects the 37kDa subunit of mouse caspase-9 only after cleavage at aspartic acid 353. Caspase-12 Antibody detects full-length mouse caspase-12 protein (55 kDa) and its cleaved product (42 kDa). Cleaved PARP (Asp214) (7C9) mouse mAb (Mouse Specific) detects the large fragment (89 kDa) of mouse PARP1 resulting from caspase cleavage.

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to amino-terminal residues adjacent to Asp175 of human caspase-3 protein, residues surrounding Asp387 of mouse caspase-8 protein, or carboxy-terminal residues surrounding Asp214 in mouse PARP protein. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to a region within the large 18 kDa subunit of mouse caspase-8 protein, residues adjacent to Asp353 of mouse caspase-9 protein, or residues surrounding amino acid 158 of mouse caspase-12 protein. 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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