Product Name
Caspase-9 Recombinant Rabbit Monoclonal Antibody [SZ29-01] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Caspase-9 aa 289-338 / 416.
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Target Molecular Weight
Predicted band size: 46 kDa
Positive Control
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.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
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.
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
Mitochondrion, cytoplasm, cytosol, nucleus.
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
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
Caspase 9 Dominant Negative antibody
Caspase 9c antibody
Caspase-9 antibody
Caspase-9 subunit p10 antibody
ICE LAP6 antibody
ICE like apoptotic protease 6 antibody
ICE-LAP6 antibody
ICE-like apoptotic protease 6 antibody
MCH6 antibody
PPP1R56 antibody
protein phosphatase 1, regulatory subunit 56 antibody
RNCASP9 antibody
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
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
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
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.
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
ICC staining of Caspase-9 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1603-27, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Caspase-9 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1603-27, 1/50) for 30 minutes 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.
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
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.
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon 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.
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
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.
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse spleen 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.
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This data was developed using ET1603-27, the same antibody clone in a different buffer formulation.
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).
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