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Caspase-3 p12 Recombinant Rabbit Monoclonal Antibody [JE07-51]

Usd: 385

Specification

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Catalog# HA722235

Caspase-3 p12 Recombinant Rabbit Monoclonal Antibody [JE07-51]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

Caspase-3 p12 Recombinant Rabbit Monoclonal Antibody [JE07-51]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Caspase-3 p12 aa 128-277 / 277.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 32/12 kDa

Positive Control

HeLa cell lysate, HeLa treated with 1μM staurosporine for 4 hours cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 1μM staurosporine for 3 hours cell lysate, C6 cell lysate, C6 treated with 1μM staurosporine for 4 hours cell lysate, human breast cancer tissue, human tonsil tissue, mouse liver tissue, mouse kidney tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

JE07-51

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P FC IF-Cell IP
human
Tested Tested
Tested Tested
Not Recommended
Not Recommended
Not Recommended
mouse
Tested Tested
Tested Tested
 
 
 
rat
Tested Tested
Tested Tested
 
 
 

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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

  • IHC-P

  • 1:200-1:1,000

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).

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 Caspase-3 p12 on different lysates with Rabbit anti-Caspase-3 p12 antibody (<a href="/products/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</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: NIH/3T3 cell lysate<br />Lane 4: NIH/3T3 treated with 1μM staurosporine for 3 hours cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: C6 treated with 1μM staurosporine for 4 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 32/12 kDa<br />Observed band size: 32/12 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<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/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</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 Caspase-3 p12 on different lysates with Rabbit anti-Caspase-3 p12 antibody (HA722235) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 1μM staurosporine for 4 hours cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: NIH/3T3 treated with 1μM staurosporine for 3 hours cell lysate
    Lane 5: C6 cell lysate
    Lane 6: C6 treated with 1μM staurosporine for 4 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 32/12 kDa
    Observed band size: 32/12 kDa

    Exposure time: 3 minutes; ECL: K1802;
    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 (HA722235) 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;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Caspase-3 p12 on different lysates with Rabbit anti-Caspase-3 p12 antibody (<a href="/products/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/2,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate (10 µg/Lane)<br />Lane 2: HAP1-Caspase-3 p12 KD cell lysate (10 µg/Lane)<br /><br />Predicted band size: 32 kDa<br />Observed band size: 32 kDa<br /><br />Exposure time: 40 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/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/2,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> 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.

    ☑ Knockdown (KD)

    Western blot analysis of Caspase-3 p12 on different lysates with Rabbit anti-Caspase-3 p12 antibody (HA722235) at 1/2,000 dilution.

    Lane 1: HAP1-parental cell lysate (10 µg/Lane)
    Lane 2: HAP1-Caspase-3 p12 KD cell lysate (10 µg/Lane)

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

    Exposure time: 40 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 (HA722235) at 1/2,000 dilution was used in K1803 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 breast cancer tissue with Rabbit anti-Caspase-3 p12 antibody (<a href="/products/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/200 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/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/200 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 breast cancer tissue with Rabbit anti-Caspase-3 p12 antibody (HA722235) at 1/200 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 (HA722235) at 1/200 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 tonsil tissue with Rabbit anti-Caspase-3 p12 antibody (<a href="/products/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/200 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/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/200 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 tonsil tissue with Rabbit anti-Caspase-3 p12 antibody (HA722235) at 1/200 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 (HA722235) at 1/200 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 mouse liver tissue with Rabbit anti-Caspase-3 p12 antibody (<a href="/products/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/200 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/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</a>) at 1/200 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 mouse liver tissue with Rabbit anti-Caspase-3 p12 antibody (HA722235) at 1/200 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 (HA722235) at 1/200 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 mouse kidney tissue with Rabbit anti-Caspase-3 p12 antibody (<a href="/products/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</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/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</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 mouse kidney tissue with Rabbit anti-Caspase-3 p12 antibody (HA722235) 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 (HA722235) 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 rat kidney tissue with Rabbit anti-Caspase-3 p12 antibody (<a href="/products/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</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/HA722235" style="font-weight: bold;text-decoration: underline;">HA722235</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 rat kidney tissue with Rabbit anti-Caspase-3 p12 antibody (HA722235) 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 (HA722235) 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.

Citation

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