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Pro Caspase-3 Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER30804

Pro Caspase-3 Rabbit Polyclonal Antibody

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

  • IHC-Fr

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Pro Caspase-3 Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within N-terminal human Caspase-3.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell, IHC-P, FC, IHC-Fr

Molecular Weight

Predicted band size: 32 kDa

Positive Control

Jurkat cell lysate, Ramos cell lysate, human brain tissue lysate, HepG2, A549, human liver carcinoma tissue, human tonsil tissue.

Conjugation

unconjugated

RRID

AB_3069554

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IF-Cell

  • 1:200

  • IHC-P

  • 1:200

  • FC

  • 1:100-1:200

  • IHC-Fr

  • 1:100

Target

Function

Caspase-3 is a caspase protein that interacts with caspase-8 and caspase-9. It is encoded by the CASP3 gene. CASP3 orthologs have been identified in numerous mammals for which complete genome data are available. Unique orthologs are also present in birds, lizards, lissamphibians, and teleosts. Caspase-3 shares many of the typical characteristics common to all currently-known caspases. For example, its active site contains a cysteine residue (Cys-163) and histidine residue (His-121) that stabilize the peptide bond cleavage of a protein sequence to the carboxy-terminal side of an aspartic acid when it is part of a particular 4-amino acid sequence. This specificity allows caspases to be incredibly selective, with a 20,000-fold preference for aspartic acid over glutamic acid. A key feature of caspases in the cell is that they are present as zymogens, termed procaspases, which are inactive until a biochemical change causes their activation. Each procaspase has an N-terminal large subunit of about 20 kDa followed by a smaller subunit of about 10 kDa, called p20 and p10, respectively.

Background References

1. "In vitro activation of CPP32 and Mch3 by Mch4, a novel human apoptotic cysteine protease containing two FADD-like domains." Fernandes-Alnemri T., Armstrong R.C., Krebs J.F., Srinivasula S.M., Wang L., Bullrich F., Fritz L.C., Proc. Natl. Acad. Sci. U.S.A. 93:7464-7469(1996)

2. "Potent and selective nonpeptide inhibitors of caspases 3 and 7 inhibit apoptosis and maintain cell functionality." Lee D., Long S.A., Adams J.L., Chan G., Vaidya K.S., Francis T.A., Kikly K., Winkler J.D., Sung C.-M., Nuttall M.E.J. Biol. Chem. 275:16007-16014(2000)

Sequence Similarity

Belongs to the peptidase C14A family.

Tissue Specificity

Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

Post-translational Modification

Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.; S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.

Subcellular Location

Nucleus, cytoplasm,

UNIPROT

SwissProt: P42574 Human

Synonyms

Caspase3

A830040C14Rik antibody

Apopain antibody

CASP 3 antibody

CASP-3 antibody

CASP3 antibody

CASP3_HUMAN antibody

Casp3a antibody

Caspase 3 antibody

Caspase 3, apoptosis-related cysteine peptidase antibody

Expand

Images

  • Western blot analysis of Pro Caspase-3 on different lysates with Rabbit anti-Pro Caspase-3 antibody (<a href="/products/ER30804" style="font-weight: bold;text-decoration: underline;">ER30804</a>) at 1/5,000 dilution.<br /><br />Lane 1: Jurkat cell lysate<br />Lane 2: Ramos cell lysate<br />Lane 3: Human brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 32 kDa<br />Observed band size: 32 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER30804" style="font-weight: bold;text-decoration: underline;">ER30804</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Pro Caspase-3 on different lysates with Rabbit anti-Pro Caspase-3 antibody (ER30804) at 1/5,000 dilution.

    Lane 1: Jurkat cell lysate
    Lane 2: Ramos cell lysate
    Lane 3: Human brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 32 kDa
    Observed band size: 32 kDa

    Exposure time: 3 minutes; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER30804) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining of Caspase-3 in HepG2 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining of Caspase-3 in HepG2 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • ICC staining of Caspase-3 in A549 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

    ICC staining of Caspase-3 in A549 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

  • Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Caspase-3 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Caspase-3 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Caspase-3 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Caspase-3 antibody. Counter stained with hematoxylin.

  • Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Pro Caspase-3 with Rabbit anti-Pro Caspase-3 antibody (<a href="/products/ER30804" style="font-weight: bold;text-decoration: underline;">ER30804</a>).<br />The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ER30804" style="font-weight: bold;text-decoration: underline;">ER30804</a>, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor&reg; 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

    Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Pro Caspase-3 with Rabbit anti-Pro Caspase-3 antibody (ER30804).
    The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ER30804, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

  • Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Pro Caspase-3 with Rabbit anti-Pro Caspase-3 antibody (<a href="/products/ER30804" style="font-weight: bold;text-decoration: underline;">ER30804</a>).<br />The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ER30804" style="font-weight: bold;text-decoration: underline;">ER30804</a>, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor&reg; 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

    Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Pro Caspase-3 with Rabbit anti-Pro Caspase-3 antibody (ER30804).
    The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (ER30804, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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