Caspase-3 Antibody (CPP32 4-1-18) - BSA Free

Western Blot: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - ALM induces HIF-1alpha-dependent apoptosis in PC3 cells. PC3 cells were treated with ALM for 24 h and 48 h at 0.1 uM under normoxia and hypoxia, followed by Western blot for caspase-3. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2073-4409/8/5/439), licensed under a CC-BY license.

Western Blot: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - ALM induces HIF-1alpha-dependent apoptosis in PC3 cells. PC3 cells were treated with ALM for 24 h and 48 h at 0.1 uM under normoxia and hypoxia, followed by Western blot for caspase-3. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2073-4409/8/5/439), licensed under a CC-BY license.

Simple Western: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Lane view shows a specific band for Caspase 3 in 0.5 mg/ml of Hek293 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Analysis of a FFPE human spleen section using 1:200 dilution of . The staining was developed using HRP conjugated anti-mouse secondary antibody and DAB reagent. This Caspase 3 antibody generated a specific staining in the cytoplasm of various spleenocytes.

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Caspase-3 was detected in immersion fixed paraffin-embedded sections of human bladder 1:300 dilution of mouse monoclonal Caspase-3 Antibody (CPP32 4-1-18) (NB500-210, Novus Biologicals), for 1 hour at room temperature followed by anti-mouse IgG VisUCyte HRP polymer(VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue).

Immunohistochemistry-Frozen: Caspase-3 Antibody (CPP32 4-1-18) - BSA Free [NB500-210] - Analysis of murine spleen with Caspase-3 (green), CD3 (red) and DAPI (blue). Image from verified customer review.

Western Blot: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Detection of Caspase (19 and 35 kDa) from HEK293 cell extract using (NB500-210). Lanes 1 and 2 contain inactive and active Caspase, respectively.

Immunohistochemistry-Paraffin: Caspase-3 Antibody (CPP32 4-1-18) [NB500-210] - Rat epithelial cells of the tongue base. Antigen retrieval method: Citrate buffer.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF, IHC, IP
• Clone: CPP32 4-1-18
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Caspase-3 Antibody (CPP32 4-1-18) - BSA Free Summary

• Immunogen: This Caspase-3 Antibody (CPP32 4-1-18) was developed against full-length recombinant human Caspase 3 [UniProt# P42574].
• Isotype: IgG2a Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: CASP3
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 2 ug/ml
  • Immunohistochemistry 1:200-1:500
  • Immunohistochemistry-Frozen reported in scientific literature (PMID 31242448)
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Immunoprecipitation 2 ug / mg lysate
  • Simple Western 1:2000
  • Western Blot 1:500-1:1000
• Application Notes: A band is seen at ~32 kDa for the inactive form of Caspase 3 and ~17-22 kDa for the active form of Caspase 3 in Western Blot. (See protocol for additional information).
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in Hek293 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:2000, apparent MW was 40 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Theoretical MW: 31.7 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  293 Whole Cell Lysate	Loading...
  Jurkat Staurosporine Treated / Untreated Cell Lysate	Loading...

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Caspase-3 Antibody (CPP32 4-1-18) - BSA Free

• Apopain
• apoptosis-related cysteine protease
• CASP3
• CASP-3
• caspase 3, apoptosis-related cysteine peptidase
• Caspase3
• Caspase-3
• CPP32
• CPP-32
• CPP32B
• CPP32SREBP cleavage activity 1
• Cysteine protease CPP32
• EC 3.4.22
• EC 3.4.22.56
• LICE-1
• PARP cleavage protease
• procaspase3
• Protein Yama
• SCA-1
• YAMA

Background

CPP32 encodes a member of the cysteine-aspartic acid protease (caspase) family that plays a key role in apoptosis. Caspase 3 (also known as CPP32, Apopain, Yama, and PARP cleavage protease) is the most extensively studied apoptotic protein among caspase family members (1-2). Caspase 3 is synthesized as an inactive proenzyme that is processed in cells undergoing apoptosis by self-proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10). Alternative splicing of CPP32 results in two transcript variants which encode the same protein. The processed form of Caspase 3 consists of large (theoretical molecular weight 17kD) and small (theoretical molecular weight 12kD) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28/Ser29 and Asp175/Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Variations in levels of Caspase 3 have been reported in cells of short-lived nature and those with a longer life cycle. Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease (3).

References

1.Mu, N., Lei, Y., Wang, Y., Wang, Y., Duan, Q., Ma, G., . . . Su, L. (2019). Inhibition of SIRT1/2 upregulates HSPA5 acetylation and induces pro-survival autophagy via ATF4-DDIT4-mTORC1 axis in human lung cancer cells. Apoptosis, 24(9-10), 798-811. doi:10.1007/s10495-019-01559-3

2.Sun, C. M., Enkhjargal, B., Reis, C., Zhou, K. R., Xie, Z. Y., Wu, L. Y., . . . Zhang, J. H. (2019). Osteopontin attenuates early brain injury through regulating autophagy-apoptosis interaction after subarachnoid hemorrhage in rats. CNS Neurosci Ther, 25(10), 1162-1172. doi:10.1111/cns.13199

3.Louneva, N., Cohen, J. W., Han, L. Y., Talbot, K., Wilson, R. S., Bennett, D. A., . . . Arnold, S. E. (2008). Caspase-3 is enriched in postsynaptic densities and increased in Alzheimer's disease. Am J Pathol, 173(5), 1488-1495. doi:10.2353/ajpath.2008.080434

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Caspase-3 Antibody (NB500-210)(24)

Publications using NB500-210

• Basavaraj P, Hsieh PF, Jiang WP et al. Elucidation of scandenolone as anti-cancer activity through impairment of the metabolic and signaling vulnerabilities in prostate cancer Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 2023-05-29 [PMID: 37257224] (WB, Human)
• Liu Z, Wang M, Huang R et al. Novel Indole-Chalcone Derivative-Ligated Platinum(IV) Prodrugs Attenuate Cisplatin Resistance in Lung Cancer through ROS/ER Stress and Mitochondrial Dysfunction Journal of medicinal chemistry 2023-03-22 [PMID: 36946996]
• Hsieh P, Jiang W, Basavaraj P Et al. Cell suspension culture extract of Eriobotrya japonica attenuates growth and induces apoptosis in prostate cancer cells via targeting SREBP-1/FASN-driven metabolism and AR Phytomedicine 2021-12-01 [PMID: 34740154] (WB, Human)
• Rutledge EA, Parvez RK, Short KM et al. Morphogenesis of the kidney and lung requires branch-tip directed activity of the Adamts18 metalloprotease Dev. Biol. 2019-06-23 [PMID: 31242448] (IHC-F, Mouse)
• Cheng Jiadong, Hu Lan, Yang Zheng et al. 2-Oxonanonoidal Antibiotic Actinolactomycin Inhibits Cancer Progression by Suppressing HIF-1 alpha. Cells 2019-05-10 [PMID: 31083403] (WB, Human)
• Martins FF, Aguila MB, Mandarim-de-Lacerda CA. Impaired steroidogenesis in the testis of leptin-deficient mice (ob/ob -/-). Acta Histochem. 2017-05-13 [PMID: 28506466]
• Graus-Nunes F, Marinho TS, Barbosa-da-Silva S et al. Differential effects of angiotensin receptor blockers on pancreatic islet remodelling and glucose homeostasis in diet-induced obese mice. Mol. Cell. Endocrinol. 2016-10-22 [PMID: 27780713] (IF/IHC, Mouse)
• Kim H, Lee KH, Park IA et al. Expression of SIRT1 and apoptosis-related proteins is predictive for lymph node metastasis and disease-free survival in luminal A breast cancer. Virchows Arch. 2015-08-18 [PMID: 26280894] (IHC-P, Human)
  
  Details:
  Caspase 3 antibody was used at 1:300 dilution for IHC-P analysis of tissue sections from human cases of luminal A invasive breast ductal carcinoma. The assay was performed on benchmark automatic immunostaining device and the signal detection was performed using biotinylated anti-mouse secondary antibody -peroxidase-labeled streptavidin - DAB method.
• Shibayama Y, Kondo T, Ohya H et al. Upregulation of microRNA-126-5p is associated with drug resistance to cytarabine and poor prognosis in AML patients. Oncol Rep 2015-05-01 [PMID: 25759982]
• Hsuan SW, Chyau CC, Hung HY et al. The induction of apoptosis and autophagy by Wasabia japonica extract in colon cancer Eur J Nutr 2015-02-27 [PMID: 25720497] (WB, Human)
• Wu SH, Chyau CC, Chen JH et al. Anti-cancerous effects of Wasabia japonica extract in Hep3B liver cancer cells via ROS accumulation, DNA damage and p73-mediated apoptosis Journal of Functional Foods 2015-03-02 (WB, Human)
• Su C Sann-Joong-Kuey-Jian-Tang decreases the protein expression of mammalian target of rapamycin but increases microtubule associated protein II light chain 3 expression to inhibit human BxPC 3 pancreatic carcinoma cells. Mol Med Rep 2015-01-04 [PMID: 25516264] (WB, Human)
• Hu H, Takano N, Xiang L et al. Hypoxia-inducible factors enhance glutamate signaling in cancer cells. Oncotarget 2014-10-15 [PMID: 25326682] (WB, Human)
• Chien SY, Kuo SJ, Chen DR, Su CC. Sann-Joong-Kuey-Jian-Tang decreases the protein expression of Mcl?1 and TCTP and increases that of TNF-alpha and Bax in BxPC?3 pancreatic carcinoma cells. Int J Mol Med 2013-07-01 [PMID: 23652631]
• Chen YL, Yan MY, Chien SY et al. Sann-Joong-Kuey-Jian-Tang inhibits hepatocellular carcinoma Hep-G2 cell proliferation by increasing TNF-alpha, Caspase-8, Caspase- 3 and Bax but by decreasing TCTP and Mcl-1 expression in vitro. Mol Med Rep 2013-05-01 [PMID: 23525225] (Human)
• Peng Z, Shuangzhu Y, Yongjie J et al. TNF receptor-associated factor 6 regulates proliferation, apoptosis, and invasion of glioma cells. Mol Cell Biochem 2013-01-29 [PMID: 23358926] (WB, Human)
• Fuzio P, Ditonno P, Rutigliano M et al. Regulation of TGF-B1 expression by Androgen Deprivation Therapy of prostate cancer Cancer Letters 2012-05-18 [PMID: 22269108] (IF/IHC, Human)
• Hsu L-C, Hsu Y-W, Liang Y-H et al. Induction of Apoptosis in Human Breast Adenocarcinoma Cells MCF-7 by Monapurpyridine A, a New Azaphilone Derivative from Monascus purpureus NTU 568. Molecules. 2012-01-01 [PMID: 22237681]
• Dash R, Azab B, Quinn B et al. Apogossypol derivative BI-97C1 (Sabutoclax) targeting Mcl-1 sensitizes prostate cancer cells to mda-7/IL-24-mediated toxicity Proc Natl Acad Sci U S A 2011-05-24 [PMID: 21555592] (WB, Human)
• Hsu LC, Hsu YW, Liang YH, Kuo YH, Pan TM. Anti-tumor and anti-inflammatory properties of ankaflavin and monaphilone A from monascus purpureus NTU 568. J Agric Food Chem;59(4):1124-30. 2011-02-23 [PMID: 21261255] (WB, Mouse)
• Rothberg BEG, Bracken MB, Rimm DL. Tissue Biomarkers for Prognosis in Cutaneous Melanoma: A Systematic Review Meta-analysis. J Natl Cancer Inst;101(7):452-474. 2009-01-01 [PMID: 19318635]
• Kulkarni AA, Kingsbury SR, Tudzarova S et al. Cdc7 kinase is a predictor of survival and a novel therapeutic target in epithelial ovarian carcinoma. Clin Cancer Res 2009-04-01 [PMID: 19318489]
• Nikiforov MA et al. Tumor cell-selective regulation of NOXA by c-MYC in response to proteasome inhibition. Proc Natl Acad Sci U S A104(49):19488-93. 2007-12-04 [PMID: 18042711]
• Rodriguez J, Lazebnik Y. Caspase-9 and APAF-1 form an active holoenzyme. Genes Dev;13(24):3179-84. 1999-12-15 [PMID: 10617566]

Reviews for Caspase-3 Antibody (NB500-210) (4)

Average Rating: 4

(Based on 4 reviews)

We have 4 reviews tested in 2 species: Human, Mouse.

reviewed by: Verified Customer IHC-Fr Mouse 12/05/2022

Summary

• Application: Immunohistochemistry-Frozen
• Sample Tested: Mouse Spleen
• Species: Mouse
• Lot: C7

reviewed by: Verified Customer WB Human 08/23/2015

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell whole cell lysate
• Species: Human

reviewed by: Verified Customer WB Human 12/21/2011

Summary

• Application: Western Blot
• Sample Tested: Human Hepatoblastoma
• Species: Human

reviewed by: Verified Customer WB Human 11/21/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human

FAQs for Caspase-3 Antibody (NB500-210). (Showing 1 - 2 of 2 FAQs).

1. I am looking for a mouse antibody against active caspase 3. This antibody you have is one of the few mouse anti-caspase 3 antibodies that I can find, but the information about it does not specify whether it is against the active form of the protein. I must have an antibody against active caspase 3, can you tell me whether this is the case for your antibody (NB500-210).  ..show answer..
   • I have pulled up information about this antibody and it appears that it will detect both the active and non-active forms of the protein. I don't know if you are looking for one that is only specific to the active and will not detect the inactive, or if as long as it detects the active it will work.
2. Which is the best retrieval buffer for this antibody to IHC? In addition, which is the dilution for it?  ..show answer..
   • Antigen retrieval should be performed using citrate buffer at a pH of 6.0. Our protocol for this can be found here: Antigen Retrieval Protocol.

Bioinformatics

• Gene Symbol: CASP3
• Entrez:
  • 836(Human)
  • 12367(Mouse)
  • 25402(Rat)
• Uniprot:
  • P42574(Mouse)