Caspase-3 Antibody (31A893)

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.

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 anti-Caspase 3 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. Right panel: Jurkat cells were treated with 5uM Camptothecin for 4 hours, 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 anti-Caspase 3 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.

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 anti-Caspase-3 monoclonal (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).

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

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

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

Caspase-3 Antibody (31A893) Summary

• Immunogen: A recombinant full-length human Caspase-3 protein was used as the immunogen for this antibody.
• Isotype: IgG
• Clonality: Monoclonal
• Host: Mouse
• Gene: CASP3
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/mL
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 5 ug/mL
  • Immunohistochemistry-Paraffin 5 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).
• Control:

  Caspase-3 Knockout HeLa 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)

• 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

This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family and is a downstream effector cysteine protease in the apoptotic pathway. Caspases are a family of cysteine proteases that are key mediators of programmed cell death or apoptosis. 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. 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. The protein is ubiquitously expressed in normal human tissues including the liver, spleen, heart, liver and kidney.

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 ()