Name: Apoptosis Antibody Sampler Kit
Brand: Cell Signaling Technology
Price: 592.00 USD
Availability: InStock

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Apoptosis Antibody Sampler Kit: Image 1 — 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.

Apoptosis Antibody Sampler Kit: Image 2 — 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.

Apoptosis Antibody Sampler Kit: Image 3 — 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).

Apoptosis Antibody Sampler Kit: Image 4 — 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).

Apoptosis Antibody Sampler Kit: Image 5 — 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.

Apoptosis Antibody Sampler Kit: Image 6 — 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.

Apoptosis Antibody Sampler Kit: Image 7 — 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).

Apoptosis Antibody Sampler Kit: Image 8 — 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.

Apoptosis Antibody Sampler Kit: Image 9 — 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.

Apoptosis Antibody Sampler Kit: Image 10 — 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.

Apoptosis Antibody Sampler Kit: Image 11 — 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.

Apoptosis Antibody Sampler Kit: Image 12 — 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).

Apoptosis Antibody Sampler Kit: Image 13 — Immunohistochemical analysis of paraffin-embedded human tonsil using Cleaved PARP (Asp214) (D64E10) XP^® Rabbit mAb.

Apoptosis Antibody Sampler Kit: Image 14 — 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).

Apoptosis Antibody Sampler Kit: Image 15 — 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.

Apoptosis Antibody Sampler Kit: Image 16 — 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).

Apoptosis Antibody Sampler Kit: Image 17 — Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved Caspase-7 (Asp198) (D6H1) Rabbit mAb.

Apoptosis Antibody Sampler Kit: Image 18 — 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.

Apoptosis Antibody Sampler Kit: Image 19 — 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).

Apoptosis Antibody Sampler Kit: 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)).

Apoptosis Antibody Sampler Kit: Image 21 — Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

Apoptosis Antibody Sampler Kit: Image 22 — 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).

Apoptosis Antibody Sampler Kit: Image 23 — 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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To Purchase # 9915

Cat. #	Size	Qty.	Price
9915T	1 Kit (8 x 20 microliters)		$ 592

Product Includes
Related Products

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
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

Kit Usage Information

Protocols

5625: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence*, Flow
7074: Western Blotting
7076: Western Blotting
8438: Western Blotting, Immunofluorescence, Flow
9508: Western Blotting
9542: Western Blotting
9664: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
12827: Western Blotting
14220: Western Blotting, Immunoprecipitation (Agarose)
52873: Western Blotting, Immunofluorescence

Product Description

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.

Specificity / Sensitivity

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 (D3R6Y) 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.

Source / Purification

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

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.

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Apoptosis Antibody Sampler Kit #9915