Caspase-3 Antibody - (Pro and Active)

Western Blot: Caspase-3 Antibody - (Pro and Active) - Unpurified [NB100-56112] - Caspase-3 Antibody - (Pro and Active) [NB100-56112] - Analysis of Caspase-3. Lysates from Jurkat cells (lane 1), normal mammary tissue (lane 2) and surgical specimens from three invasive ductal carcinomas (lanes 3-5) were normalized for total protein content (50 ug/lane) and western blotted with anti-Caspase-3. The ~32 kDa pro-Caspase-3 protein was detected in all samples. Active/cleaved Caspase-3 was identified in Jurkat and two ductal carcinomas (14-21 kDa large subunit).

Immunohistochemistry-Paraffin: Caspase-3 Antibody - (Pro and Active) [NB100-56112] - Caspase-3 was detected in immersion fixed paraffin-embedded sections of human bladder tissue 1:300 dilution of rabbit anti-Caspase-3 polyclonal (NB100-56112, Novus Biologicals), for 1 hour at room temperature followed by anti-rabbit IgG VisUCyte HRP polymer(VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue).

Immunohistochemistry-Paraffin: Caspase-3 Antibody - (Pro and Active) [NB100-56112] - Caspase-3 expression in formalin-fixed, paraffin-embedded human reactive lymph node using NB100-56112 at 1:2000. Staining is seen in the apoptosis-prone germinal center B lymphocytes of follicles.

Immunohistochemistry-Paraffin: Caspase-3 Antibody - (Pro and Active) [NB100-56112] - Analysis of Caspase-3 expression in formalin-fixed, paraffin-embedded human breast ductal carcinoma in situ using this antibody at 1:2000. Staining is seen in the the cancerous ducts, but not in the normal lobulus.

Simple Western: Caspase-3 Antibody - (Pro and Active) - Unpurified [NB100-56112] - Caspase-3 Antibody - (Pro and Active) [NB100-56112] - Simple Western lane view shows a specific band for Caspase 3 in 0.2 mg/ml of HEK293 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Ca, Ge
• Applications: WB, Simple Western, ChIP, IHC, IHC-Fr, IHC-P, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

Caspase-3 Antibody - (Pro and Active) Summary

• Immunogen: Recombinant full-length human Caspase-3 protein (pro-form) was used as immunogen.
• Localization: The subcellular location is primarly in the cytoplasm.
• Specificity: Caspase antibodies are classical tools for detecting inactive (pro) and active (cleaved) forms of the enzymes. The presence of the large or small subunits in western blots is considered to be a marker of caspase activation.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CASP3
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:1000-1:2000
  • Simple Western 1:200
  • Chromatin Immunoprecipitation 1:10-1:500
  • Immunohistochemistry 1:1000-1:5000
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:1000-1:5000
  • Immunoprecipitation 1:50-1:200
• Application Notes: Immunoprecipitation, Western Blot, Immunohistochemistry-Paraffin IHC (frozen): Users should optimize according to model and immunodetection system used (secondary reagents). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. Use in chromatin immunoprecipitation reported in scientific literature (PMID 27735949).
• Control:

  Caspase-3 Overexpression Lysate
  Caspase-3 Knockout HeLa Cell Lysate

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Whole antisera
• Preservative: 0.05% Sodium Azide
• Purity: Unpurified

Alternate Names for Caspase-3 Antibody - (Pro and Active)

• 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

Apoptosis, or programmed cell death, is a common property of all multicellular organisms. The current dogma of apoptosis suggests that the components of the core cell-death machinery are integral to cells and widely conserved across species. Caspases, a family of cysteinyl aspartate-specific proteases, are integral components of the cell death machinery (reviewed in Siegal, 2006; and Lavrik et al, 2005). They play a central role in the initiation and execution of apoptotic cell death and in inflammation. Caspases are typically divided into 3 major groups, depending on the structure of their prodomain and their function. Group 1: inflammatory caspases (caspases 1, 4, 5, 11, 12, 14). Group II: initiator of apoptosis caspases (caspases 2, 8, 9). Group II: effector caspases (caspases 3, 6, 7). Caspases are synthesized as zymogens (inactive pro enzyme precursors which require a biochemical change to become active enzymes) with an N-terminal prodomain of variable length followed by a large subunit (p20) and a small subunit (p10). Caspases are activated through proteolytic cleavage at specific asparagine residues that are located within the prodomain, the p10, and p20 subunits. Activation results in the generation of mature active caspases that consist of the heterotetramer p202-p102. Active caspases mediate cell death and inflammation through cleavage of particular cellular substrates that are involved in these processes. The Caspase-3 polyclonal antisera recognizes the proform of caspase-3 (approx. 32) kDa), and the large (approx. 14-21 kDa) and small (approx. 10 kDa) subunits of active/cleaved Caspase-3.

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 (NB100-56112)(12)

Publications using NB100-56112

• Kwon I, Song W, Jang Y et al. Elevation of hepatic autophagy and antioxidative capacity by endurance exercise is associated with suppression of apoptosis in mice Annals of Hepatology [PMID: 31611063] (WB)
• Bean GR, Kremer JC, Prudner BC et al. A metabolic synthetic lethal strategy with arginine deprivation and chloroquine leads to cell death in ASS1-deficient sarcomas. Cell Death Dis Oct 13 2016 [PMID: 27735949] (ChIP, WB, Human)
• Cheng Tc, Lai Cs, Chung Mc et al. Potent anti-cancer effect of 3'-hydroxypterostilbene in human colon xenograft tumors PLoS OnE et al. 2014 Nov 13 [PMID: 25389774] (WB, Human)
  
  Details:
  Caspase-3 antibody used in WB for the detection of pro and cleaved forms of Caspase 3 protein in lysates of COLO 205 xenografts in nude mice treated or not with Pterostilbene and 3'-hydroxypterostilbene (Figure 6D).
• Samaga KKL, Rao GV, Chandrashekara Reddy G. Synthetic racemates of abyssinone I and II induces apoptosis through mitochondrial pathway in human cervix carcinoma cells. Bioorganic Chemistry. 2014 Jun 26 [PMID: 25019692] (WB, Human)
  
  Details:
  Fig 4: HeLa
• Krajewska Maryla, You Zerong, Rong Juan et al. Neuronal deletion of caspase 8 protects against brain injury in mouse models of controlled cortical impact and kainic acid-induced excitotoxicity. PLoS One. 2011 [PMID: 21957448] (WB, Mouse)
• Liu Q, Zhao X, Frissora F et al. FTY720 demonstrates promising preclinical activity for chronic lymphocytic leukemia and lymphoblastic leukemia/lymphoma. Blood. 2008 Jan 1 [PMID: 17761520]
• Zhao X, Lapalombella R, Joshi T et al. Targeting CD37-positive lymphoid malignancies with a novel engineered small modular immunopharmaceutical. Blood. 2007 Oct 1 [PMID: 17440052]
• Krajewska M, Rosenthal RE, Mikolajczyk J et al. Early processing of Bid and caspase-6, -8, -10, -14 in the canine brain during cardiac arrest and resuscitation. Exp Neurol. 2004 Oct [PMID: 15380478] (WB, IHC-P, Mouse, Canine, Rat)
  
  Details:
  WB and IHC (P) on mouse spleen and thymus lysates, dog and rat brain tissue and lysates, tissue culture transfectants, caspase-9 knockout mice, recombinant caspase proteins.
• Knoblach SM, Nikolaeva M, Huang X et al. Multiple caspases are activated after traumatic brain injury: evidence for involvement in functional outcome. J Neurotrauma. 2002 Oct [PMID: 12427325]
• Berger T, Kretzler M. Interaction of DAP3 and FADD only after cellular disruption. Nat Immunol. 2002 Jan [PMID: 11753396]
• Svingen PA, Karp JE, Krajewski S et al. Evaluation of Apaf-1 and procaspases-2, -3, -7, -8, and -9 as potential prognostic markers in acute leukemia. Blood. 2000 Dec 1 [PMID: 11090079]
• Krajewski S, Gascoyne RD, Zapata JM et al. Immunolocalization of the ICE/Ced-3-family protease, CPP32 (Caspase-3), in non-Hodgkin's lymphomas, chronic lymphocytic leukemias, and reactive lymph nodes. Blood. 1997 May 15 [PMID: 9160689]

Review for Caspase-3 Antibody (NB100-56112) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Anonymous WB Mouse 01/12/2017

Summary

• Application: Western Blot
• Sample Tested: BRAIN(Substantia Nigra par compacta)
• Species: Mouse
• Lot: AR14-01040-07

Bioinformatics

• Gene Symbol: CASP3