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Caspase-9 Recombinant Rabbit Monoclonal Antibody [SZ29-01]

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Catalog# ET1603-27

Caspase-9 Recombinant Rabbit Monoclonal Antibody [SZ29-01]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

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

概述

产品名称

Caspase-9 Recombinant Rabbit Monoclonal Antibody [SZ29-01]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within Human Caspase-9 aa 289-338 / 416.

种属反应性

Human, Mouse

验证应用

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

靶点分子量

Predicted band size: 46 kDa

阳性对照

C2C12 cell lysate, NIH/3T3, HepG2 cell, A549 cell, human tonsil tissue, human cervix carcinoma tissue, human colon tissue, human colon carcinoma tissue, mouse spleen tissue.

偶联

unconjugated

克隆号

SZ29-01

RRID

AB_3069677

产品特性

形态

Liquid

浓度

存放说明

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

存储缓冲液

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

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:500-1:5,000

  • IF-Cell

  • 1:100-1:500

  • IF-Tissue

  • 1:100-1:500

  • IHC-P

  • 1:400-1:5,000

  • IP

  • Use at an assay dependent concentration.

  • FC

  • 1:1,000

靶点

功能

Caspase-9 is an enzyme that in humans is encoded by the CASP9 gene. It is an initiator caspase, critical to the apoptotic pathway found in many tissues. Caspase-9 homologs have been identified in all mammals for which they are known to exist, such as Mus musculus and Pan troglodytes. 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.

背景文献

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).

序列相似性

Belongs to the peptidase C14A family.

组织特异性

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.

翻译后修饰

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.

亚细胞定位

Mitochondrion, cytoplasm, cytosol, nucleus.

UNIPROT

SwissProt: P55211 Human

SwissProt: Q8C3Q9 Mouse

别名

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

展开

图片

  • Western blot analysis of Caspase-9 on different lysates with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 (Mouse fibroblast) cell lysates <br />Lane 2: C2C12 (Mouse myoblast) cell lysates<br /><br />Lysates/proteins at 20 µg/Lane.<br />Exposure time: 24 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 46 kDa<br />Observed band size: 50 kDa

    Western blot analysis of Caspase-9 on different lysates with Rabbit anti-Caspase-9 antibody (ET1603-27) at 1/1,000 dilution.

    Lane 1: NIH/3T3 (Mouse fibroblast) cell lysates
    Lane 2: C2C12 (Mouse myoblast) cell lysates

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 24 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1603-27, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

  • Immunocytochemistry analysis of NIH/3T3 cells labeling Caspase-9 with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling Caspase-9 with Rabbit anti-Caspase-9 antibody (ET1603-27) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Caspase-9 antibody (ET1603-27) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human cervix carcinoma tissue with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/400 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/400 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 cervix carcinoma tissue with Rabbit anti-Caspase-9 antibody (ET1603-27) at 1/400 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1603-27) at 1/400 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 colon tissue with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/5,000 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 colon tissue with Rabbit anti-Caspase-9 antibody (ET1603-27) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1603-27) at 1/5,000 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 colon carcinoma tissue with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/400 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/400 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 colon carcinoma tissue with Rabbit anti-Caspase-9 antibody (ET1603-27) at 1/400 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1603-27) at 1/400 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 spleen tissue with Rabbit anti-Caspase-9 antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>) at 1/5,000 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 spleen tissue with Rabbit anti-Caspase-9 antibody (ET1603-27) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET1603-27) at 1/5,000 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.

  • Flow cytometric analysis of NIH/3T3 cells labeling Caspase-9.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1603-27" style="font-weight: bold;text-decoration: underline;">ET1603-27</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of NIH/3T3 cells labeling Caspase-9.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1603-27, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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

引文

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