Product Pathways - Apoptosis
Effector Caspases and Substrates Antibody Sampler Kit #12690
|12690S||1 Kit (6 x 40 µl)||---||In Stock||---|
Already purchased this product? Write a Review.
|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Caspase-3 (8G10) Rabbit mAb #9665||40 µl||W, IP||H, M, R, Mk||17, 19, 35||Rabbit IgG|
|Caspase-6 Antibody #9762||40 µl||W||H, M, R||15, 35||Rabbit|
|Caspase-7 (D2Q3L) Rabbit mAb #12827||40 µl||W||H, M, R||Mk||20, 35||Rabbit IgG|
|Lamin A/C (4C11) Mouse mAb #4777||40 µl||W, IP, IHC-P, IF-F, IF-IC, F||H, M, R, Mk||74 (Lamin A), 63 (Lamin C)||Mouse IgG2a|
|Lamin B1 (D4Q4Z) Rabbit mAb #12586||40 µl||W||H, M, R||Mk, Pg||68, 25||Rabbit IgG|
|PARP Antibody #9542||40 µl||W||H, M, R, Mk||89, 116||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-mouse IgG, HRP-linked Antibody #7076||100 µl||Horse|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-F=Immunofluorescence (Frozen), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +), using Lamin B1 (D4Q4Z) Rabbit mAb #12586.
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 #12827.
Western blot analysis of extracts from THP-1 cells, untreated (-) or cycloheximide-treated (CHX, 10 μg/ml, overnight; +) followed by treatment with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (20 ng/ml, 4 hr; +), using Lamin A/C (4C11) Mouse mAb #4777.
Western blot analysis of extracts from NIH/3T3 cells, untreated (-) or treated with Staurosporine #9953 (1 µM; +), and Jurkat cells, untreated (-) or treated with Etoposide #2200 (25 µM; +), using PARP Antibody #9542.
Western analysis of extracts from HeLa and NIH/3T3 cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +), using Caspase-3 (8G10) Rabbit mAb #9665.
Western blot analysis of extracts from NIH/3T3 cells, untreated (-) or treated with Staurosporine #9953 (1 µM; +) and Jurkat cells, untreated (-) or treated with Etoposide #2200 (25 µM; +), using Caspase-6 Antibody #9762.
The Effector Caspases and Substrates Antibody Sampler Kit provides an economical means to evaluate the activation of effector (executioner) caspases. The kit contains enough primary antibody to perform at least four western blots per primary antibody.
Specificity / Sensitivity
Each antibody in the Effector Caspases and Substrates Antibody Sampler Kit recognizes endogenous levels of its respective target. Caspase-3 (8G10) Rabbit mAb recognizes full-length (35 kDa) and the large fragment (17/19 kDa) of caspase-3 resulting from cleavage at Asp175. Caspase-6 Antibody recognizes full length (35 kDa) and the small subunit (15 kDa) of caspase-6 resulting from cleavage at Asp193. Caspase-7 (D2Q3L) Rabbit mAb recognizes full-length (35 kDa) and the large subunit (20 kDa) of caspase-7 resulting from cleavage at Asp198. PARP Antibody recognizes full length (116 kDa) and the large fragment (89 kDa) of PARP1 resulting from caspase cleavage at Asp214. Lamin A/C (4C11) Mouse mAb recognizes full-length lamin A and lamin C proteins, and the larger fragments of lamin A (50 kDa) and lamin C (41 kDa) resulting from caspase cleavage. Lamin B1 (D4Q4Z) Rabbit mAb recognizes endogenous levels of total lamin B1 protein and a 25 kDa fragment resulting from caspase cleavage.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues adjacent to Asp175 in human caspase-3 protein, residues surrounding Pro158 of human caspase-7 protein, residues surrounding Leu118 of human lamin B1 protein, or with a recombinant fragment of human lamin A protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the cleavage site of caspase-6 or the caspase cleavage site in PARP. 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. Caspase-3 (CPP-32, Apoptain, Yama, SCA-1), Caspase-6 (Mch2), and Caspase-7 (CMH-1, Mch3, ICE-LAP3) are effector caspases functioning in cellular apoptotic processes (1-6). Upon apoptotic stimulation, initiator caspases such as caspase-9 (ICE-LAP6, Mch6) are cleaved and activated (7). The activated upstream caspases further process downstream executioner caspases by cleaving them into activated large and small subunits, thereby initiating a caspase cascade leading to apoptosis (4,6,8-10).
PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (11). This protein can be cleaved by many ICE-like caspases in vitro (1,12) and is one of the main cleavage targets of caspase-3 in vivo (10,13). In human PARP, cleavage occurs between Asp214 and Gly215, which separates the PARP amino-terminal DNA binding domain (24 kDa) from the carboxy-terminal catalytic domain (89 kDa) (10,12). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (14).
Lamins are nuclear membrane structural components that are important in maintaining normal cell functions, such as cell cycle control, DNA replication, and chromatin organization (15-17). Lamins have been subdivided into types A and B. Type-A lamins consist of lamin A and C, which arise from alternative splicing of the lamin A gene LMNA. Lamin A and C are cleaved by caspases into large (41-50 kDa) and small (28 kDa) fragments, which can be used as markers for apoptosis (18,19). Type-B lamins consist of lamin B1 and B2, encoded by separate genes (20-22). Lamin B1 is also cleaved by caspases during apoptosis (23).
- Cohen, G.M. (1997) Biochem J 326 ( Pt 1), 1-16.
- Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4.
- Faleiro, L. et al. (1997) EMBO J 16, 2271-81.
- Fernandes-Alnemri, T. et al. (1995) Cancer Res 55, 6045-52.
- Duan, H. et al. (1996) J Biol Chem 271, 1621-5.
- Lippke, J.A. et al. (1996) J Biol Chem 271, 1825-8.
- Li, P. et al. (1997) Cell 91, 479-89.
- Slee, E.A. et al. (1999) J Cell Biol 144, 281-92.
- MacFarlane, M. et al. (1997) J Cell Biol 137, 469-79.
- Nicholson, D.W. et al. (1995) Nature 376, 37-43.
- Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-8.
- Lazebnik, Y.A. et al. (1994) Nature 371, 346-7.
- Tewari, M. et al. (1995) Cell 81, 801-9.
- Oliver, F.J. et al. (1998) J Biol Chem 273, 33533-9.
- Dunbar, J.C. and Lu, H. (2000) Brain Res Bull 52, 123-6.
- Goldberg, M. et al. (1999) Crit Rev Eukaryot Gene Expr 9, 285-93.
- Yabuki, M. et al. (1999) Physiol Chem Phys Med NMR 31, 77-84.
- Rao, L. et al. (1996) J Cell Biol 135, 1441-55.
- Orth, K. et al. (1996) J Biol Chem 271, 16443-6.
- Biamonti, G. et al. (1992) Mol Cell Biol 12, 3499-506.
- Lin, F. and Worman, H.J. (1995) Genomics 27, 230-6.
- Pollard, K.M. et al. (1990) Mol Cell Biol 10, 2164-75.
- Chandler, J.M. et al. (1997) Biochem J 322 ( Pt 1), 19-23.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7072 Phototope®-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 9998 BSA
- 9999 Nonfat Dry Milk
- 9997 Tris Buffered Saline with Tween® 20 (TBST-10X)
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7722 Blue Loading Buffer Pack
- 7723 Red Loading Buffer Pack
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7075 Anti-biotin, HRP-linked Antibody
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.