Skip to content

Cleaved+pro Caspase-3 Recombinant Rabbit Monoclonal Antibody [SU38-04]

Usd: 385

Specification

Bulk Order Quote | Customization Request

Catalog# ET1608-64

Cleaved+pro Caspase-3 Recombinant Rabbit Monoclonal Antibody [SU38-04]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

Cleaved+pro Caspase-3 Recombinant Rabbit Monoclonal Antibody [SU38-04]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Caspase-3 aa 9-175 / 277.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 32/17 kDa

Positive Control

HeLa cell lysate, HeLa treated with 1μM staurosporine for 4 hours cell lysate, HeLa treated with 3μM staurosporine for 4 hours cell lysate, C6 cell lysate, C6 treated with 1μM staurosporine for 4 hours cell lysate, C6 treated with 3μM staurosporine for 4 hours cell lysate, U-87 MG cell lysate, MDA-MB-231 cell lysate, NIH/3T3 cell lysate, Jurkat cell lysate, Jurkat treated with 1μM staurosporine for 3 hours cell lysate, human lung carcinoma tissue, human spleen tissue, human liver tissue, human kidney tissue.

Conjugation

unconjugated

Clone Number

SU38-04

RRID

AB_3069820

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:50-1:1,000

  • IP

  • Use at an assay dependent concentration.

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. Dong L et al. Echinacoside Induces Apoptosis in Human SW480 Colorectal Cancer Cells by Induction of Oxidative DNA Damages. Int J Mol Sci 16:14655-68 (2015).

2. Nilsonne G et al. Phenotype-dependent apoptosis signalling in mesothelioma cells after selenite exposure. J Exp Clin Cancer Res 28:92 (2009).

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

Cytoplasm.

UNIPROT

SwissProt: P42574 Human

SwissProt: P70677 Mouse

SwissProt: P55213 Rat

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

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Cleaved+pro Caspase-3 on different lysates with Rabbit anti-Cleaved+pro Caspase-3 antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 1μM staurosporine for 4 hours cell lysate<br />Lane 3: HeLa treated with 3μM staurosporine for 4 hours cell lysate<br />Lane 4: C6 cell lysate<br />Lane 5: C6 treated with 1μM staurosporine for 4 hours cell lysate<br />Lane 6: C6 treated with 3μM staurosporine for 4 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 32/17 kDa<br />Observed band size: 32/17 kDa<br /><br />Exposure time: 3 seconds; ECL: K1801;<br /><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/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1/1,000 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.

    ☑ Cell treatment (CT)

    Western blot analysis of Cleaved+pro Caspase-3 on different lysates with Rabbit anti-Cleaved+pro Caspase-3 antibody (ET1608-64) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 1μM staurosporine for 4 hours cell lysate
    Lane 3: HeLa treated with 3μM staurosporine for 4 hours cell lysate
    Lane 4: C6 cell lysate
    Lane 5: C6 treated with 1μM staurosporine for 4 hours cell lysate
    Lane 6: C6 treated with 3μM staurosporine for 4 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 32/17 kDa
    Observed band size: 32/17 kDa

    Exposure time: 3 seconds; 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 (ET1608-64) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Knockout (KO)</span><br /><br />Western blot analysis of Caspase-3 with anti-Caspase-3 antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1:2,000 dilution.<br />Lane 1: Wild-type Hela whole cell lysate (10 µg).<br />Lane 2: Caspase-3 knockout Hela whole cell lysate (10 µg).<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>, 1/2,000) was used in 5% BSA at room temperature for 2 hours. 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.

    ☑ Knockout (KO)

    Western blot analysis of Caspase-3 with anti-Caspase-3 antibody (ET1608-64) at 1:2,000 dilution.
    Lane 1: Wild-type Hela whole cell lysate (10 µg).
    Lane 2: Caspase-3 knockout Hela whole cell lysate (10 µg).

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1608-64, 1/2,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Cleaved+pro Caspase-3 on different lysates with Rabbit anti-Cleaved+pro Caspase-3 antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1/5,000 dilution.<br /><br />Lane 1: U-87 MG cell lysate<br />Lane 2: MDA-MB-231 cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: Jurkat cell lysate<br />Lane 5: Jurkat treated with 1μM staurosporine for 3 hours cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 32/17 kDa<br />Observed band size: 32/17 kDa<br /><br />Exposure time: 1 minute 2 seconds;<br /><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/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</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.

    ☑ Cell treatment (CT)

    Western blot analysis of Cleaved+pro Caspase-3 on different lysates with Rabbit anti-Cleaved+pro Caspase-3 antibody (ET1608-64) at 1/5,000 dilution.

    Lane 1: U-87 MG cell lysate
    Lane 2: MDA-MB-231 cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: Jurkat cell lysate
    Lane 5: Jurkat treated with 1μM staurosporine for 3 hours cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 32/17 kDa
    Observed band size: 32/17 kDa

    Exposure time: 1 minute 2 seconds;

    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 (ET1608-64) 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.

  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-Cleaved+pro Caspase-3 antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-Cleaved+pro Caspase-3 antibody (ET1608-64) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-64) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Cleaved+pro Caspase-3 antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1608-64" style="font-weight: bold;text-decoration: underline;">ET1608-64</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Cleaved+pro Caspase-3 antibody (ET1608-64) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1608-64) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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

Citation

Products with the same target and pathway

metafield image

Cleaved+pro Caspase-3 Recombinant Rabbit Monoclonal Antibody [SU38-04] - BSA and Azide free

Application: WB,IHC-P,IP

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated

metafield image

Cleaved+pro Caspase-3 Recombinant Rabbit Monoclonal Antibody [JE75-05]

Application: WB

Reactivity: Human,Mouse

Conjugate: unconjugated