Active+Pro Caspase-9 Recombinant Rabbit Monoclonal Antibody [SC65-05] - BSA and Azide free
Catalog# HA750238
Active+Pro Caspase-9 Recombinant Rabbit Monoclonal Antibody [SC65-05] - BSA and Azide free
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WB
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IP
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Human
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Mouse
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ET1610-95
含抗保成分
-
unconjugated
Overview
Product Name
Active+Pro Caspase-9 Recombinant Rabbit Monoclonal Antibody [SC65-05] - BSA and Azide free
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
Product Features
Form
Liquid
Concentration
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IP
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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.
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
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of Active+Pro Caspase-9 on different lysates with Rabbit anti-Active+Pro Caspase-9 antibody (HA750238) 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 (HA750238) 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 (HA750238) 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 (HA750238) 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. -
☑ Knockdown (KD)
Western blot analysis of Caspase-9 on different lysates with Rabbit anti-Caspase-9 antibody (HA750238) 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"
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Conjugate: unconjugated
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