Background
Caspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis (1). The precursor form of all caspases is composed of a prodomain, and large and small catalytic subunits. The active forms of caspases are generated by several stimuli including ligand-receptor interactions, growth factor deprivation and inhibitors of cellular functions. All known caspases require cleavage adjacent to aspartates to liberate one large and one small subunit, which associate into a₂b₂ tetramer to form the active enzyme. Gene for Caspase-3 also known as Yama, CPP32, and apopain codes for a 32-kDa protein (2-4).  Caspase-3 cleaves the death substrate poly(ADP-ribose) polymerase (PARP) to a specific 85 kDa form observed during apoptosis and is inhibitable by the CrmA protein. Other Caspase-3 substrates include DNA-PK, actin, GAS2, and procaspase-6, etc. (5). Caspase-3 is activated by cleavage events at Asp-28/Ser-29 (between N-terminal pro-domain) and Asp-175/Ser-176 (between large and small subunits) to generate a large subunit of 17-kDa and a small subunit of 12-kDa (3).

Antigen
Full-length recombinant human Caspase-3 protein was used as immunogen. 

The antibody recognizes an epitope in the large domain subunit of Caspase-3.  As such it will recognize pro Caspase-3 and the large subunit cleavage fragment.

Application Notes
The antibody detects both pro Caspase-3 (~32 kDa) and the large subunit of the active/cleaved form (~14-21 kDa) of Caspase-3.  The large subunit of the cleaved form may appear as one or two or even as a stack of bands depending on the presence or absence of the Caspase-3 pro-domain.

Genebank Info (Protein)
NP_033940

Related Products

Reference
1. Cohen GM. Biochem J. 326:1-16, 1997.
2. Fernandes-Almnemri T, et al. J. Biol. Chem. 269:30761-4, 1994.
3. Tewari M, et al. Cell 81: 801-809, 1995.
4. Shi L, et al. Proc. Natl. Acad. Sci. USA 93:11002-7, 1996.
5. P, et al. TIBS 22: 389-392, 1997.

Product Citations
1. Apoptotic Proteins in the Temporal Cortex in Schizophrenia: High Bax/Bcl-2 Ratio Without Caspase-3 Activation.  L. Fredrik Jarskog, Elzbieta S. Selinger, Jeffrey A. Lieberman, and John H. Gilmore.  Am. J. Psychiatry, Jan 2004; 161: 109 - 115. IMGENEX antibodies cited: 1. Caspase-3 (IMG-144) [WB, Fig 1 ( Postmortem Brain Tissue)] 
2. Ricin Triggers Apoptotic Morphological Changes through Caspase-3 Cleavage of BAT3. Wu, Y-H, et al. J. Biol. Chem., 279: 19264-19275 (2004). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144) [WB, Fig.3C (HeLa cells)].
3. Molecular Cloning and Characterization of Bif-1. Cuddeback S. M., H. Yamaguchi, K. Komatsu, T. Miyashita, M. Yamada, C. Wu, S. Singh, H. G. Wang. Biol Chem, 276: 20559-20565 (2001). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [ WB, Fig 3C, (FL5.12 cells) ] 2. Bif-1 (IMG-265A) [ WB, Fig 8A, (FL5.12 cells) ].
4. Epothilone B Analogue (BMS-247550)-mediated Cytotoxicity through Induction of Bax Conformational Change in Human Breast Cancer Cells. Hirohito Yamaguchi, Shanthi R. Paranawithana, Michael W. Lee, Ziwei Huang, Kapil N. Bhalla, and Hong-Gang Wang. Cancer Res., 62: 466-471 (2002). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 1D (468 cells) ]. 2. D4-GDI (IMG-160A) [WB, Fig 1D (468 cells) ]. 
5. The Synthetic Retinoid CD437 Selectively Induces Apoptosis in Human Lung Cancer Cells while Sparing Normal Human Lung Epithelial Cells. Shi-Yong Sun, Ping Yue, Xueyun Chen, Waun K. Hong, and Reuben Lotan. Cancer Res., 62: 2430 – 2436 (2002). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144) [WB, Fig 4A,B (H460 human lung cancer cell lines, and NHBE normal lung bronchus cells) ]
6. The protein kinase PKB/Akt regulates cell survival and apoptosis by inhibiting Bax conformational change. Hirohito Yamaguchi, Hong-Gang Wang. Oncogene  20, 7779-7786 (2001). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [ WB, Fig 2B (FL5.12 cells) ]. 2. D4-GDI (IMG-160A) [WB, Fig 2B (FL5.12 cells) ].
7. Identification of a Novel Synthetic Triterpenoid, Methyl-2-cyano-3,12-dioxooleana-1,9-dien-28-oate, That Potently Induces Caspase-mediated Apoptosis in Human Lung Cancer Cells. Kevin B. Kim, Reuben Lotan, Ping Yue, Michael B. Sporn, Nanjoo Suh, Gordon W. Gribble, Tadashi Honda, Gen Sheng Wu, Waun Ki Hong, and Shi-Yong Sun. Mol. Cancer Ther., 1: 177-184 (2002). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 4C, 4D (H460 and A549 cells) ].  
8. Identification of Retinamides That Are More Potent than N-(4-Hydroxyphenyl)retinamide in Inhibiting Growth and Inducing Apoptosis of Human Head and Neck and Lung Cancer Cells. Shi-Yong Sun, Ping Yue, Gary J. Kelloff, Vernon E. Steele, Scott M. Lippman, Waun K. Hong, and Reuben Lotan. Cancer Epidemiol. Biomarkers Prev., 10: 595-601 (2001). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 4 AB (H292 cells) ].
9. Emodin induces apoptosis in human promyeloleukemic HL-60 cells accompanied by activation of caspase 3 cascade but independent of reactive oxygen species production.  Yen-Chou Chen, Shing-Chuan Shen, Woan-Ruoh Lee, Foun-Lin Hsu, Hui-Yi Lin, Ching-Huai Ko and Shi-Wen Tseng. Biochemical Pharmacology  64, 1713-1724 (2002). IMGENEX antibodies cited: 1. Caspace-3 (IMG-144A) [WB, Fig 4A,  Fig 6A, Fig 8B (HL-60 cells) ] 2. PARP (IMG-401A) [WB, Fig 4A,  Fig 6A, Fig 8B (HL-60 cells) ] 3. D4-GDI (IMG-160A) [WB, Fig 4A,  Fig 6A, Fig 8B (HL-60 cells) ]. 
10. Therapeutic time window of adenovirus-mediated GDNF gene transfer after transient middle cerebral artery occlusion in rat.  W. R. Zhang, K. Sato, M. Iwai, I. Nagano, Y. Manabe and K. Abe. Brain Research  947, 1140-1145 (2002). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [IHC(Frozen), Fig 2, Table 1 (rat brain) ].
11. Reversal of Taxol resistance in hepatoma by cyclosporin A: involvement of the PI-3 kinase-AKT 1 pathway. L Lin, W-Y Lui, T-Y Liu, C-W Chi. British Journal of Cancer  88, 973-980 (2003). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 4 (HepG2 cells) ].
12. Ricin Triggers Apoptotic Morphological Changes through Caspase-3 Cleavage of BAT3. Yu-Hauh Wu, Sheue-Fang Shih, and Jung-Yaw Lin . J. Biol. Chem., 279: 19264-19275 (2004). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 3B (HeLa cells) ].
13. Hydroxylation At C4v Or C6 Is Essential For Apoptosis-Inducing Activity Of Flavanone Through Activation Of The Caspase-3 Cascade And Production Of Reactive Oxygen Species. Ching Huai Ko,Shing-Chuan Shen,Y,Z And Yen Chou Chen. Free Radical Biology & Medicine, 36 (7): 897–910 (2004). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) 2. PARP (IMG-80405) 3. D4-GDI (IMG-396A). [WB, Fig 4AB, 5C, 7A, 8D, 9C, 10 (HL-60 human promyeloleukemic cells) ].
14. Lipolysis Products from Triglyceride-Rich Lipoproteins Increase Endothelial Permeability, Perturb Zonula Occludens-1, F-actin, and Induce Apoptosis. Larissa Eiselein, Dennis Wilson, Michael Lame, and John C. Rutledge. Am J Physiol Heart Circ Physiol, Jan 2007; 10.1152/ajpheart.00686.2006. IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 5B (T-75 cells) ].
15. Synergism of Rana catesbeiana ribonuclease and IFN-a triggers distinct death machineries in different human cancer cells. Chih-Hang Anthony Tanga, Chih-Chi Andrew Huc, Chyou-Wei Weia, and Jaang-Jiun Wang. FEBS Lett. 579 (1): 265-270 (2005). WB; MCF-7, SK-Hep-1, HL-60 cells treated with RC-RNase, Fig 5.
16. Overexpression of Bcl-2 is associated with apoptotic resistance to the G-quadruplex ligand 12459 but is not sufficient to confer resistance to long-term senescence. Céline Douarre, Dennis Gomez, Hamid Morjani, Jean-Marie Zahm, Marie-Françoise O'Donohue, Lahcen Eddabra, Patrick Mailliet, Jean-François Riou, and Chantal Trentesaux. Nucleic Acids Res  33: 2192-2203 (2005).
1. WB, A549 (human lung carcinoma) cells treated with triazine derivative 12549, Fig 4C.
2. WB, untreated A549 , JFD18, JFD9 cells, Fig 3b.
17. Justicidin A decreases the level of cytosolic Ku70 leading to apoptosis in human colorectal cancer cells. Jenq-Chang Lee, Chao-Hung Lee, Chun-Li Su, Chung-Wei Huang, Hsiao-Sheng Liu, Chun-Nan Lin, and Shen-Jeu Won. Carcinogenesis, May 2005; 10.1093/carcin/bgi133. IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 3A (HT-29 and HCT 116 cells) ].  
18. Activation of Nuclear Factor-kB Contributes to Induction of Death Receptors and Apoptosis by the Synthetic Retinoid CD437 in DU145 Human Prostate Cancer Cells. Fengshuo Jin, Xiangguo Liu, Zhongmei Zhou, Ping Yue, Reuben Lotan, Fadlo R. Khuri, Leland W.K. Chung, and Shi-Yong Sun. Cancer Res.,  65: 6354-6363 (2005).
19. Induction of endoplasmic reticulum stress and apoptosis by a marine prostanoid in human hepatocellular carcinoma. Po-Cheng Chiang, Chung-Liang Chien, Shiow-Lin Pan, Wen-Pin Chen, Che-Ming Teng, Ya-Ching Shen and Jih-Hwa Guh. Journal of Hepatology, 43 (4): 679-686 (2005).
20. Induction of endothelial cell apoptosis by lipid hydroperoxide-derived bifunctional electrophiles. Wenying Jian, Jasbir S. Arora, Tomoyuki Oe, Vladimir V. Shuvaev and Ian A. Blair. Free Radical Biology and Medicine, 39 (9): 1162-1176 (2005). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 1BD, Fig 3C, Fig 7ABC, Fig 8ABC, (EA.hy 926 endothelial cells) ].
21. YC-1 suppresses constitutive nuclear factor-kB activation and induces apoptosis in human prostate cancer cells. Yao-Ting Huang, Shiow-Lin Pan, Jih-Hwa Guh, Ya-Ling Chang, Fang-Yu Lee, Sheng-Chu Kuo, and Che-Ming Teng. Mol. Cancer Ther., 4: 1628-1635 (2005). IMGENEX antibodies cited: 1. Caspase-3 (IMG-144A) [WB, Fig 5A (human prostate cancer PC-3 cells) ].
22. Caspase-2 triggers Bax-Bak dependent and independent cell death in colon cancer cells treated with resveratrol. John Mohan, Alankaram Arul Gandhi, Balan Chandrika Bhavya, Ramachandran Rashmi, Devarajan Karunagaran, Ramachandran Indu, and Thankayyan Retnabhai Santhoshkumar. J. Biol. Chem., Apr 2006; 10.1074/jbc.M602641200. IMGENEX products cited: 1. Caspase-3 (IMG-144) [WB, Fig 1C,  Fig 7C, Fig 8 (human colon adenocarcinoma cell line, HCT 116) ]  2.  AIF (IMG-303) [WB, Fig 2B, IF/FCC, Fig 2B  (HCT 116 cells) ]  3. RNAi vector (IMG-805) [ Fig 3CD (HCT 116 cells) ].
23. Induction of Fas clustering and apoptosis by coral prostanoid in human hormone-resistant prostate cancer cells. Po-Cheng Chiang, Fan-Lu Kung, Dong-Ming Huang, Tsai-Kun Li, Jia-Rong Fan, Shiow-Lin Pan, Ya-Ching Shen and Jih-Hwa Guh. European Journal of Pharmacology, In Press, Accepted Manuscript, Available online 3 June 2006. IMGENEX antibodies cited:  1. Caspase-3 (IMG-144A) [WB, Fig 4, 6B (human hormone-resistant prostate cancer PC-3 cells) ]. 
24. Caspase-3 activation in rat frontal cortex following treatment with typical and atypical antipsychotics. Jarskog FL, JH Gilmore, LA Glantz, KL Gable, TT German, RI Tong, JA Lieberman. Neuropsychopharmacology 32:95-102 (2007). Rat frontal cortex tissue: WB (Figs 1a and b, Table 1), IHC (frozen), Fig 3a and b.  
25. Troglitazone sensitizes tumor cells to TRAIL-induced apoptosis via down-regulation of FLIP and Survivin. Kerstin Schultze, Barbara Böck, Anika Eckert, Lena Oevermann, Dirk Ramacher, Otmar Wiestler and Wilfried Roth. Apoptosis 11: 1503-1512 (2006).
WB, human glioma and neuroblastoma cells treated with either TRAIL or troglitazone, Fig 2D.
26. The Immediate Early 2 Protein of Human Cytomegalovirus (HCMV) Mediates the Apoptotic Control in HCMV Retinitis through Up-Regulation of the Cellular FLICE-Inhibitory Protein Expression. Shih-Hwa Chiou, Yi-Ping Yang, Jung-Chun Lin, Chih-Hung Hsu, Hua-Ci Jhang, Yu-Ting Yang, et al. J Immunol., 177: 6199-6206 (2006).
WB, mock or HCMV-infected HRPE cells, Fig 4B.
27. The alkylphospholipid perifosine induces apoptosis of human lung cancer cells requiring inhibition of Akt and activation of the extrinsic apoptotic pathway. Heath A. Elrod, Yi-Dan Lin, Ping Yue, Xuerong Wang, Sagar Lonial, Fadlo R. Khuri, and Shi-Yong Sun.Mol. Cancer Ther., Jun 2007; 10.1158/1535-7163.MCT-07-0004. IMGENEX antibodies cited: Caspase-3 (IMG-144A) [WB, Fig 2BC,  Fig 4B, Fig 5B (H157 and H460 cells) ].
28. Protein expression profiling and molecular clasification of gastric cancer by the tissue array method.  Lee H S, S-B Cho, HE Lee, MA Kim, JH Kim, DJ Park, JH Kim, H-K Yang, BL Lee, WH Kim.  Clinical Cancer Research 13: 4154-4163 (2007).  Imgenex products cited:
1. Gastric Carcinoma Tissue Microarray (IMH-316): IHC (paraffin), Tables 1,2; Fig.1.
2. Stomach Tissue Microarray (IMH-341), matching normal tissue of IMH-316: IHC (paraffin), Tables 1,2; Fig. 1 
3. Caspase-3 (IMG-144A): IHC (paraffin) on gastric carcinoma microarrays, Tables 1,2.  In nonneoplastic mucosa faint diffuse caspase-3 staining was observed. In gastric cancer Caspase-3 was expressed definitely in the nucleus and cytoplasm. 
4. Rad9 (IMG-164): IHC (paraffin) on gastric carcinoma microarrays, Tables 1,2; Fig.1. In nonneoplastic mucosa Rad9 was expressed in the nucleus.  In gastric cancer there was a loss of Rad9 expression.
29. CHM-1, a novel synthetic quinolone with potent and selective antimitotic antitumor activity against human hepatocellular carcinoma in vitro and invivo. Wang S, S Pan, Y Huang, J Guh, P Chiang, D Huang, S Kuo, K Lee, C Teng. Mol Cancer Ther 7: 350-360 (2008). Imgenex antibodies cited:
1. IMG-144A: WB (Human heptatocellular carcinoma cell line HA22T), Fig. 4A.
30. Deoxycholic acid can induce apoptosis in the human colon cancer cell line HCT116 in the absence of bax. Yui S, R Kanamoto, T Saeki. Nutrition and Cancer 60: 91-96 (2008). IMG-144A: WB (human  HCT116 cells), Fig. 3C.
31. BLOC1S2 interacts with the HIPP1 protein and sensitizes NCH89 glioblastoma cells to apoptosis.  Gdynia G, J Lehmann-Kock, S Sieber, K Tagscherer, A Fassl, H Zentgraf, S Matzuzawa, J Reed, W Roth. Apoptosis 13: 437-447 (2008).  WB, human glioblastoma cells, Fig 6A.
32. Chondroitin-4-sulphate reduced oxidative injury in caerulein-induced pancreatitis in mice: the involvement of NF-_kB translocation and apoptosis activation.  Campo G, A Anvenso, S Campo, G Nastasi, P Traina, A D'Ascola, A Calatroni. Experimental Biology and Medicine 233: 741-752 (2008). Imgenex antbodies cited:
1. Goat Anti-Rabbit HRP Conjugate (20301):  ELISA (mouse pancreactic tissue), Fig. 1B
2. Caspase-3 (IMG-144A): WB (mouse pancreatic tissue), Fig. 4B
3. Caspase-7 (IMG-5560): WB (mouse pancreatic tissue), Fig. 5B.
33. Monitoring caspase-3 activation with a multimodality imaging sensor in living subjects. Ray P, A De, M Patel, S Gambhir. Clinical Cancer Research 14: 5801-5809 (2008). WB (human 293T cells), Fig. 2D.
34. Monitoring caspase-3 activation with a multimodaliyt imaging sensor in living subjects. Ray P, A De, M Patel, S Gambhir. Clinical Cancer Research 14: 5801-5809 (2008). WB (human 293T cells), Fig. 2D,E.  293T cells were transfected with CMV-mtf plasmid.
35. Comparison of the cardiac effects between quinazoline-based α₁-adrenoceptor antagonists on occlusion-reperfusion injury. Lee A, M Su. Journal of Biomedical Science 15:239-249 (2008). WB(rat heart tissue lysates), Fig. 4.
36. BLOC1S2 interacts with the HIPPI protein and sensitizes NCH89 glioblastoma cells to apoptosis. Gdynia G, J Lehmann-Koch, S Sieber, K Tagscherer, A Fassl, H Zentgraf, S matsuzawa, J Reed W Roth. Apoptosis: 10.1007/s10495-007-0176-3 (2008). WB (human glioblastoma cell line NCH89), Fig. 6A.
37. Mana-Hox displays anticancer activity against prostate cancer cells through tubulin depolymerization and DNA damage stress. Hsiao C, Y Ho, J Hsu , W Chang, Y Chen, Y Shen, P Lyu, J Guh. Naunyn-Schmiedeberg's Archives of Pharmacology doi:10.1007/s00210-007-0330-7 (2008). WB (human prostate cancer cell line PC-3), Fig. 5.
38. Denbinobin induces apoptosis by apoptosis inducing factor releasing and DNA damage in human colorectal cancer HCT-116 cells. Chen T, S Pan, J Guh, C Chen, Y Huang, H Pai, C Teng. Naunyn-Schmiedeberg's Archives of Pharmacology doi: 10.1007/s00210-008-0324-5 (2008). WB (human colorectal cancer cell lines HCT-116 and HT-29), Fig. 3c.
39. CAAT/Enhancer binding protein homologous protein-dependent death receptor 5 induction is a major component of SHetA2-induced apoptosis in lung cancer. Lin Y, S Chen, P Yue, W Zou, D Benbrook, S Liu, T Le, K Berlin, F Khuri, S Sun. Cancer Research 68: 5335-5344 (2008). Imgenex antibodies cited [WB ( human non-small cell lung cancer cell lines)]:
1. Anti-caspase-3 (IMG-144A): Figs. 1A,B; 5A.
2. DR4 (NT) TRAIL-R1 (IMG-141A): Figs. 3A.. .
40. Coupling of endoplasmic reticulum stress to CDDO-Me-Induced up-regulation of death receptor 5 via a CHOP-dependent Mechanism involving JNK activation. Zou W, P Yue, F Khuri, S Sun. Cancer Research 68: 7484-7492 (2008). WB (human H1792 cells), Fig. 4B.
41. The natural product honokoil preferentially inhibits cellular FLICE-inhibitory protein and augments death receptor induced apoptosis. Raja S, S Chen, P Yue, T Acker, B Lefkove, J Arbiser, F Khuri, S Sun. Molecular Cancer Therapeutics 7: 2212-2223 (2008). Fig. 2C.
42. Moscatilin induces apoptosis in human colorectal cancer cells: a crucial role of c-Jun NH_2-terminal protein kinase activation caused by tubulin depolymerization and DNA damage. Chen T, S Pan, J Guh, G Liao, D Huang, C Chen, C Teng. Clinical Cancer Research 14: 4250-4258 (2008). IMG-144A: WB (human colorectal cancer HCT-116 cells), Fig. 4A,B.
43. Lipopolysaccharide extends the lifespan of mouse primary-cultured microglia. Kaneko YS, A Nakashima, K Mori, T Nagatsu, and A Ota. Brain Res 1279:9-20 (2009). WB (primary cultures of mouse microglia from the brains of newborn C57BL/6 mice), Fig 4A.  A band corresponding to the ~17 kDa fragment of active caspase-3 is seen in the UV-treated cultures along with an ~32 kDa pro-caspase band.    
44. CHM-1, a new vascular targeting agent, induces apoptosis of human umbilical vein endothelial cells via p53-mediated death receptor 5 up-regulation. Tsai A, S Pan, H Sun, C Wang, C Peng, S Wang, Y Chang, S Kuo, K Lee, C Teng. J Bio Chem 285: 5497-5506 (2010). Caspase-3 (IMG-144A): WB (human umbilical vein endothelial cells), Fig 4A, Fig 4B, Fig 5D, Fig 6B.