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9930
Apoptosis Antibody Sampler Kit (Mouse Preferred)
Primary Antibodies
Antibody Sampler Kit

Apoptosis Antibody Sampler Kit (Mouse Preferred) #9930

Citations (18)
Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells treated with Cytochrome C using Caspase-3 (D3R6Y) Rabbit mAb #14220. 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 293T cells, mock transfected (-) or transfected with a construct expressing N-terminal Myc/DDK-tagged full-length mouse Caspase-12 (Myc/DDK-mCasp12; +) using Caspase-12 (E9T3W) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from HC11 Mammary Epithelium cells, untreated (-) or treated in vitro with cytochrome c (250 μg/mL, 45 min; +) and dATP (2 mM, 45 min; +), using Caspase-12 (E9T3W) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
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 various cell lines, untreated (-) or treated with Staurosporine #9953 (1 μM; 3 hr) or with Etoposide #2200 (25 μM, overnight), using Caspase-3 (D3R6Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). MCF7 cells are negative for caspase-3 expression.
Western blot analysis of extracts from BaF3, Raw264.7 and C2C12 cell lines, using Caspase-8 Antibody.
Western blot analysis of extracts from varioius cells using Caspase-12 (E9T3W) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of caspase-12 in J774A.1 cells is consistent with the predicted expression pattern.
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.
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 293T cells, mock transfected (-) or transfected with a construct overexpressing mouse caspase-8 (mCasp8) (+), using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb.
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.
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).
Western blot analysis of NIH/3T3 cells, untreated or staurosporine-treated (1 µM), using Cleaved PARP (Asp214) (7C9) Mouse mAb.
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 HCT116 cells (lane 1) or CASP3 knock-out cells (lane 2) using Caspase-3 (D3R6Y) Rabbit mAb #14220 (upper), and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The absence of signal in the CASP3 knock-out HCT116 cells confirms specificity of the antibody for CASP3.
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.
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 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 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of NIH/3T3 cells, staurosporine-treated (left) or untreated (right), using Cleaved Caspase-9 (Asp353) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of A20 cells, untreated (blue) or treated with etoposide #2200 (25 uM, overnight; green) using Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb 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 # 9930
Cat. # Size Qty. Price
9930T
1 Kit  (8 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Caspase-3 (D3R6Y) Rabbit mAb 14220 20 µl
  • WB
  • IP
H M R Mk 35, 19, 17 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 4927 20 µl
  • WB
M 45, 57 Rabbit 
Cleaved Caspase-8 (Asp387) (D5B2) XP® Rabbit mAb 8592 20 µl
  • WB
  • IP
  • IF
  • F
M 18, 43 Rabbit IgG
Caspase-9 Antibody 9504 20 µl
  • WB
M 37, 39, 49 Rabbit 
Cleaved Caspase-9 (Asp353) Antibody 9509 20 µl
  • WB
  • IF
M 37 Rabbit 
Cleaved PARP (Asp214) (7C9) Mouse mAb 9548 20 µl
  • WB
M 89 Mouse IgG2b
Caspase-12 (E9T3W) Rabbit mAb 58208 20 µl
  • WB
M 55, 38, 28 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
Horse 

Product Description

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.

Specificity / Sensitivity

Each antibody in the Apoptosis Antibody Sampler Kit (Mouse Preferred) recognizes endogenous levels of its respective target. Caspase-3 (D3R6Y) Rabbit mAb detects full-length caspase-3 (35 kDa) as well as the large subunit (17/19 kDa) of caspase-3 resulting from cleavage during apoptosis. 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. Non-specific proteins that are induced by apoptosis under certain conditions may be detected. Caspase-12 (E9T3W) Rabbit mAb detects full-length mouse caspase-12 protein (55 kDa) and the large subunit of Caspase-12 associated with activation. Cleaved PARP (Asp214) (7C9) mouse mAb detects the large fragment (89 kDa) of PARP1 resulting from caspase cleavage.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp387 of mouse caspase-8 protein, or carboxy-terminal residues surrounding Asp214 in mouse PARP protein. Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the p20 subunit of human caspase-3 protein, or recombinant protein specific to a region within the large subunit of mouse Caspase-12 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, or residues adjacent to Asp353 of mouse caspase-9 protein. 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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