Caspase-3 Antibody (31A893) - BSA Free

Western Blot: Caspase-3 Antibody (31A893) [NB100-56709] - Curcumin Analogs Induce Apoptosis in Cancerous Cells by Several Pathways. E6-1 cells were treated for 24 hours with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK and the studied compounds, lysed and subjected to Western blot analysis. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-01230-4) licensed under a CC-BY licence.

Immunohistochemistry-Paraffin: Caspase-3 Antibody (31A893) [NB100-56709] - Tissue section of normal human breast using 5 ug/mL concentration of . Very strong diffused as well as granular immunopositivity of Caspase 3 was observed specifically in the cytoplasmic of ductal /acinar epithelial cells.

Western Blot: Caspase-3 Antibody (31A893) [NB100-56709] - Analysis for human Caspase-3 using HL60 lysates with Caspase-3 Antibody (31A893) at 2 ug/mL dilution. NB100-56709 only detects a 32 kD Caspase-3 corresponding to pro-Caspase-3.

Immunocytochemistry/Immunofluorescence: Caspase-3 Antibody (31A893) [NB100-56709] - Left panel: Untreated Jurkat cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with at 10 ug/mL overnight at 4C and detected with an anti-Mouse IgG Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: Caspase-3 Antibody (31A893) [NB100-56709] - Analysis of Caspase-3 in Jurkat cells. Cells were treated with 2 uM staurosporine for different time periods. Caspase-3 activation is detected in Western blots by the presence of Caspase-3 cleavage fragments. These antibodies detect both pro (full-length) and active (cleaved) Caspase-3, depending on the treatment time points. A basal level of endogenous active Caspase-3 may be see in untreated Jurkat cells.

Immunohistochemistry-Paraffin: Caspase-3 Antibody (31A893) [NB100-56709] - Caspase-3 was detected in immersion fixed paraffin-embedded sections of human bladder tissue using 5 ug/mL of mouse monoclonal Caspase-3 Antibody (31A893) (NB100-56709, 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).

• Reactivity: Hu
• Applications: WB, ICC/IF, IHC, IHC-P
• Clone: 31A893
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Caspase-3 Antibody (31A893) - BSA Free Summary

• Immunogen: This Caspase-3 Antibody (31A893) was developed against recombinant full-length human Caspase-3 protein.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Mouse
• Gene: CASP3
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 5 ug/mL
  • Immunohistochemistry-Paraffin 5 ug/ml
  • Western Blot 2 ug/ml
• Application Notes: For IHC-P validation of this target, 1M EDTA pH 9.0 buffer was used for antigen retrieval (citrate buffer pH 6.0 did not work).
• 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:

  Jurkat Staurosporine Treated / Untreated Cell Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for Caspase-3 Antibody (31A893) - 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 (NB100-56709)(7)

Publications using NB100-56709

• Philion C, Ma D, Ruvinov I et al. Cymbopogon citratus and Camellia sinensis extracts selectively induce apoptosis in cancer cells and reduce growth of lymphoma xenografts in vivo. Oncotarget 2017 Nov 18 [PMID: 29340014] (WB)
• Ma D, Gilbert T, Pignanelli C et al. Exploiting mitochondrial and oxidative vulnerabilities with a synthetic analog of pancratistatin in combination with piperlongumine for cancer therapy FASEB J. 2017 Sep 19 [PMID: 28928246] (WB, Human)
• Pignanelli C, Ma D, Noel M et al. Selective Targeting of Cancer Cells by Oxidative Vulnerabilities with Novel Curcumin Analogs. Sci Rep Apr 24 2017 [PMID: 28439094] (WB, Human)
• Wang WT, Chen YH, Hsu JL et al. Terfenadine induces anti-proliferative and apoptotic activities in human hormone-refractory prostate cancer through histamine receptor-independent Mcl-1 cleavage and Bak up-regulation. Naunyn Schmiedebergs Arch. Pharmacol. 2014 Jan 1 [PMID: 24048439] (WB, Human)
  
  Details:
  PC3 human prostate cancer cells, Fig 4a. Terfenadine treatment induced time-dependent caspase-3 cleavage.
• Schultz CR, Golembieski WA, King DA et al. Inhibition of HSP27 alone or in combination with pAKT inhibition as therapeutic approaches to target SPARC-induced glioma cell survival. Mol Cancer. 2012 Apr 5 [PMID: 22480225]
• Hsiao CJ, Li TK, Chan YL et al. WRC-213, an l-methionine-conjugated mitoxantrone derivative, displays anticancer activity with reduced cardiotoxicity and drug resistance: identification of topoisomerase II inhibition and apoptotic machinery in prostate cancers. Biochem Pharmacol. 2008 Feb 15 [PMID: 18035333] (WB, Human)
  
  Details:
  WB (human androgen-independent prostate cancer PC-3 cells), Fig. 5B.
• Chen JY, Chi CW, Chen HL et al. TNF-alpha renders human peritoneal mesothelial cells sensitive to anti-Fas antibody-induced apoptosis. Nephrol Dial Transplant. 2003 Sep [PMID: 12937219]

Bioinformatics

• Gene Symbol: CASP3
• Entrez:
  • 836(Human)
• Uniprot:
  • P42574()