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Active+Pro Caspase-9 Recombinant Rabbit Monoclonal Antibody [SC65-05]

Usd: 385

Specification

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Catalog# ET1610-95

Active+Pro Caspase-9 Recombinant Rabbit Monoclonal Antibody [SC65-05]

Application

  • WB

  • IP

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

Active+Pro Caspase-9 Recombinant Rabbit Monoclonal Antibody [SC65-05]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Caspase-9 aa 95-335 / 416.

Product Specificity

Caspase-9 [SC65-05] Rabbit mAb detects endogenous levels of the pro form of caspase-9 as well as cleaved fragments.

Species Reactivity

Human, Mouse

Validated Applications

WB, IP

Molecular Weight

Predicted band size: 46 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, C2C12 cell lysate, Jurkat cell lysate.

Conjugation

unconjugated

Clone Number

SC65-05

RRID

AB_2940743

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

  • 1:500-1:2,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

Caspase-9 belongs to a family of caspases, cysteine-aspartic proteases involved in apoptosis and cytokine signalling. Apoptotic signals cause the release of cytochrome c from mitochondria and activation of apaf-1 (apoptosome), which then cleaves the pro-enzyme of caspase-9 into the active dimer form. Regulation of this enzyme occurs through phosphorylation by an allosteric inhibitor, inhibiting dimerization and inducing a conformational change. Correct caspase-9 function is required for apoptosis, leading to the normal development of the central nervous system. Caspase-9 has multiple additional cellular functions that are independent of its role in apoptosis. Nonapoptotic roles of caspase-9 include regulation of necroptosis, cellular differentiation, innate immune response, sensory neuron maturation, mitochondrial homeostasis, corticospinal circuit organization, and ischemic vascular injury.

Background References

1. Arango-Gonzalez B et al. Identification of a common non-apoptotic cell death mechanism in hereditary retinal degeneration. PLoS One 9:e112142 (2014).

2. Schattenberg JM et al. Increased hepatic fibrosis and JNK2-dependent liver injury in mice exhibiting hepatocyte-specific deletion of cFLIP. Am J Physiol Gastrointest Liver Physiol 303:G498-506 (2012).

Sequence Similarity

Belongs to the peptidase C14A family.

Tissue Specificity

Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes.

Post-translational Modification

Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase-3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events.; Phosphorylated at Thr-125 by MAPK1/ERK2. Phosphorylation at Thr-125 is sufficient to block caspase-9 processing and subsequent caspase-3 activation. Phosphorylation on Tyr-153 by ABL1/c-Abl; occurs in the response of cells to DNA damage.

Subcellular Location

Apoptosome, cytosol, Mitochondrion, nucleus, cytoplasm, protein-containing complex.

UNIPROT

SwissProt: P55211 Human

SwissProt: Q8C3Q9 Mouse

Synonyms

Caspase9

APAF-3 antibody

APAF3 antibody

Apoptosis related cysteine peptidase antibody

Apoptotic protease Mch-6 antibody

Apoptotic protease-activating factor 3 antibody

CASP-9 antibody

CASP9 antibody

CASP9_HUMAN antibody

Caspase 9 apoptosis related cysteine peptidase antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Active+Pro Caspase-9 on different lysates with Rabbit anti-Active+Pro Caspase-9 antibody (<a href="/products/ET1610-95" style="font-weight: bold;text-decoration: underline;">ET1610-95</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 /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 46 kDa<br />Observed band size: 46/37/35 kDa<br /><br />Exposure time: 30 seconds; 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/ET1610-95" style="font-weight: bold;text-decoration: underline;">ET1610-95</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 Active+Pro Caspase-9 on different lysates with Rabbit anti-Active+Pro Caspase-9 antibody (ET1610-95) 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

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 46 kDa
    Observed band size: 46/37/35 kDa

    Exposure time: 30 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 (ET1610-95) 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.

  • Western blot analysis of Active+Pro Caspase-9 on different lysates with Rabbit anti-Active+Pro Caspase-9 antibody (<a href="/products/ET1610-95" style="font-weight: bold;text-decoration: underline;">ET1610-95</a>) at 1/1,000 dilution.<br /><br />Lane 1: C2C12 cell lysate<br />Lane 2: HeLa cell lysate<br />Lane 3: Jurkat cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 46 kDa<br />Observed band size: 46 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% 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/ET1610-95" style="font-weight: bold;text-decoration: underline;">ET1610-95</a>) at 1/1,000 dilution was used in 5% NFDM/TBST 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 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Active+Pro Caspase-9 on different lysates with Rabbit anti-Active+Pro Caspase-9 antibody (ET1610-95) at 1/1,000 dilution.

    Lane 1: C2C12 cell lysate
    Lane 2: HeLa cell lysate
    Lane 3: Jurkat cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 46 kDa
    Observed band size: 46 kDa

    Exposure time: 2 minutes;

    12% 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 (ET1610-95) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Caspase-9 on different lysates with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1610-95" style="font-weight: bold;text-decoration: underline;">ET1610-95</a>) at 1/1,000 dilution.<br /><br />Lane 1: Hela-si NT cell lysate<br />Lane 2: Hela-si Caspase-9 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 46 kDa<br />Observed band size: 46 kDa<br /><br />Exposure time: 2 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/ET1610-95" style="font-weight: bold;text-decoration: underline;">ET1610-95</a> was shown to specifically react with Caspase-9 in Hela-si NT cells. Weakened band was observed when Hela-si Caspase-9 sample was tested. Hela-si NT and Hela-si Caspase-9 samples were subjected to SDS-PAGE. 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/ET1610-95" style="font-weight: bold;text-decoration: underline;">ET1610-95</a>, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were 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 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Caspase-9 on different lysates with Rabbit anti-Caspase-9 antibody (ET1610-95) at 1/1,000 dilution.

    Lane 1: Hela-si NT cell lysate
    Lane 2: Hela-si Caspase-9 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 46 kDa
    Observed band size: 46 kDa

    Exposure time: 2 minutes;

    4-20% SDS-PAGE gel.

    ET1610-95 was shown to specifically react with Caspase-9 in Hela-si NT cells. Weakened band was observed when Hela-si Caspase-9 sample was tested. Hela-si NT and Hela-si Caspase-9 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-95, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

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

Citation

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