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  • Product Name
    Cleaved-Caspase 9 (Asp353) Antibody
  • Catalog No.
    AF5240
  • RRID
    AB_2837726
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Human, Mouse, Rat
  • Prediction
    Horse(100%), Dog(100%)
  • UniProt
  • Mol.Wt
    10kD,47kD;
    46kDa(Calculated).
  • Concentration
    1mg/ml
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

APAF-3; APAF3; Apoptosis related cysteine peptidase; Apoptotic protease Mch-6; Apoptotic protease-activating factor 3; CASP-9; CASP9; CASP9_HUMAN; Caspase 9 apoptosis related cysteine peptidase; Caspase 9 Dominant Negative; Caspase 9c; Caspase-9; Caspase-9 subunit p10; ICE LAP6; ICE like apoptotic protease 6; ICE-LAP6; ICE-like apoptotic protease 6; MCH6; PPP1R56; protein phosphatase 1, regulatory subunit 56; RNCASP9;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Horse(100%), Dog(100%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Specificity:

Cleaved-Caspase 9 (Asp353) Antibody detects endogenous levels of fragment of activated Caspase 9 resulting from cleavage adjacent to Asp353.

RRID:

AB_2837726
Please cite this product as: Affinity Biosciences Cat# AF5240, RRID:AB_2837726.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information

Immunogen:

A synthesized peptide derived from human Caspase 9 (Cleaved-Asp353).

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

CASP9

Molecular Weight:

Observed Mol.Wt.: 10kD,47kD.
Predicted Mol.Wt.: 46kDa(Calculated)..

Tissue Specificity:

P55211 CASP9_HUMAN:
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.

Description:

Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).

Sequence:
MDEADRRLLRRCRLRLVEELQVDQLWDALLSRELFRPHMIEDIQRAGSGSRRDQARQLIIDLETRGSQALPLFISCLEDTGQDMLASFLRTNRQAAKLSKPTLENLTPVVLRPEIRKPEVLRPETPRPVDIGSGGFGDVGALESLRGNADLAYILSMEPCGHCLIINNVNFCRESGLRTRTGSNIDCEKLRRRFSSLHFMVEVKGDLTAKKMVLALLELAQQDHGALDCCVVVILSHGCQASHLQFPGAVYGTDGCPVSVEKIVNIFNGTSCPSLGGKPKLFFIQACGGEQKDHGFEVASTSPEDESPGSNPEPDATPFQEGLRTFDQLDAISSLPTPSDIFVSYSTFPGFVSWRDPKSGSWYVETLDDIFEQWAHSEDLQSLLLRVANAVSVKGIYKQMPGCFNFLRKKLFFKTS

Research Background

Function:

Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Promotes DNA damage-induced apoptosis in a ABL1/c-Abl-dependent manner. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).

Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9.

Post-translational Modifications:

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.

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.

Subunit Structure:

Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 35 kDa (p35) and a 10 kDa (p10) subunit. Caspase-9 and APAF1 bind to each other via their respective NH2-terminal CED-3 homologous domains in the presence of cytochrome C and ATP. Interacts (inactive form) with EFHD2. Interacts with HAX1. Interacts with BIRC2/c-IAP1, XIAP/BIRC4, BIRC5/survivin, BIRC6/bruce and BIRC7/livin. Interacts with ABL1 (via SH3 domain); the interaction is direct and increases in the response of cells to genotoxic stress and ABL1/c-Abl activation. Interacts with NleF from pathogenic E.coli.

Similarity:

Belongs to the peptidase C14A family.

Research Fields

Research Fields:

· Cellular Processes > Cell growth and death > p53 signaling pathway.(View pathway)
· Cellular Processes > Cell growth and death > Apoptosis.(View pathway)
· Cellular Processes > Cell growth and death > Apoptosis - multiple species.(View pathway)
· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.(View pathway)
· Human Diseases > Cancers: Specific types > Pancreatic cancer.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Colorectal cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Endometrial cancer.(View pathway)
· Human Diseases > Neurodegenerative diseases > Parkinson's disease.
· Human Diseases > Cardiovascular diseases > Viral myocarditis.
· Human Diseases > Infectious diseases: Bacterial > Legionellosis.
· Human Diseases > Cancers: Specific types > Small cell lung cancer.(View pathway)
· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.
· Human Diseases > Infectious diseases: Viral > Influenza A.
· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.
· Human Diseases > Neurodegenerative diseases > Huntington's disease.
· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.
· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.
· Human Diseases > Cancers: Specific types > Prostate cancer.(View pathway)
· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.(View pathway)

Reference Citations:

1). Yin Z;Jiang K;Shi L;Fei J;Zheng J;Ou S;Ou J; et al. Formation of di-cysteine acrolein adduct decreases cytotoxicity of acrolein by ROS alleviation and apoptosis intervention. J Hazard Mater 2020 Apr 5;387:121686. (PubMed: 31780296) [IF=9.038]

2). Yin Z;Jiang K;Shi L;Fei J;Zheng J;Ou S;Ou J; et al. Formation of di-cysteine acrolein adduct decreases cytotoxicity of acrolein by ROS alleviation and apoptosis intervention. J Hazard Mater 2020 Apr 5;387:121686. (PubMed: 31780296) [IF=9.038]

3). Yin L et al. Bacillus spore-based oral carriers loading curcumin for the therapy of colon cancer. J Control Release 2018 Feb 10;271:31-44 (PubMed: 29274436) [IF=7.727]

4). Ding Q et al. The role of the apoptosis-related protein BCL-B in the regulation of mitophagy in hepatic stellate cells during the regression of liver fibrosis. Exp Mol Med 2019 Jan 11;51(1):6 (PubMed: 30635551) [IF=5.418]

5). Wang L et al. Development of anisamide-targeted PEGylated gold nanorods to deliver epirubicin for chemo-photothermal therapy in tumor-bearing mice. Int J Nanomedicine 2019 Mar 8;14:1817-1833 (PubMed: 30880982) [IF=5.115]

6). Yin L et al. Over-Expression of Inhibitor of Differentiation 2 Attenuates Post-Infarct Cardiac Fibrosis Through Inhibition of TGF-β1/Smad3/HIF-1α/IL-11 Signaling Pathway. Front Pharmacol 2019 Nov 13;10:1349 (PubMed: 31803053) [IF=4.225]

7). Yi J et al. Protective mechanisms of purified polyphenols from pinecones of Pinus koraiensis on spleen tissues in tumor-bearing S180 mice in vivo. Food Funct 2017 Jan 25;8(1):151-166 (PubMed: 27924972) [IF=4.171]

8). Chen F et al. A ROS-mediated lysosomal-mitochondrial pathway is induced by ginsenoside Rh2 in hepatoma HepG2 cells. Food Funct 2015 Dec;6(12):3828-37 (PubMed: 26449932) [IF=4.171]

Application: WB    Species:human;    Sample:HepG2

Fig. 5 Involvement of caspases in apoptosis induced by GRh2. (A) After incubating GRh2-loaded cells with or without 1 μM of Leu. Caspase activity was determined as indicated in section 2. Enzymatic activity is expressed as fold induction with respect to non-treated cells and represents the mean values ± SD. of three different experiments. *p < 0.001 significantly different from the control; #p < 0.001 significantly different from the cells treated with GRh2 in the absence of Leu. (B) Western blot assays of active fragments of caspase-3 and -9. The results from one representative experiment are shown. (C) The histogram represents quantification of caspase-3 and caspase-9 protein expression levels in GRh2 stimulated HepG2 cell samples using ImageJ64 software (levels of control cells/β-actin defined as 1). Results are presented as mean ± S.D. with triplicate measurement. *P < 0.01 vs. the control group.


9). Song Z et al. The protective role of the MKP-5-JNK/P38 pathway in glucolipotoxicity-induced islet β-cell dysfunction and apoptosis. Exp Cell Res 2019 Jun 13:111467 (PubMed: 31202710) [IF=3.383]

10). Luan P et al. Neuroprotective effect of salvianolate on cerebral ischaemia-reperfusion injury in rats by inhibiting the Caspase-3 signal pathway. Eur J Pharmacol 2020 Jan 21:172944 (PubMed: 31978424) [IF=3.263]

11). Shang J et al. CircPAN3 mediates drug resistance in acute myeloid leukemia through the miR-153-5p/miR-183-5p-XIAP axis. Exp Hematol 2018 Nov 2 (PubMed: 30395908)

12). Xiao Y et al. Clusterin increases mitochondrial respiratory chain complex I activity and protects against hexavalent chromium-induced cytotoxicity in L-02 hepatocytes. Toxicol Res (Camb) 2018 Nov 15;8(1):15-24 (PubMed: 30713657)

13). Wu J et al. A novel bis-aryl urea compound inhibits tumor proliferation via cathepsin D-associated apoptosis. Anticancer Drugs 2020 Jun;31(5):500-506 (PubMed: 31917700)

14). Shang J et al. CircPAN3 contributes to drug resistance in acute myeloid leukemia through regulation of autophagy. Leuk Res 2019 Aug 2;85:106198 (PubMed: 31401408)

15). Zhang X et al. Lappaconitine Sulfate Inhibits Proliferation and Induces Apoptosis in Human Hepatocellular Carcinoma HepG2 Cells through the Reactive Oxygen Species-Dependent Mitochondrial Pathway. Pharmacology 2020 Feb 14:1-10 (PubMed: 32062649)

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Catalog Number :

AF5240-BP
(Blocking peptide available as AF5240-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Horse
100%
Dog
100%
Pig
71%
Chicken
0%
Rabbit
0%
Zebrafish
0%
Xenopus
0%
Sheep
0%
Bovine
0%
High similarity Medium similarity Low similarity No similarity
P55211 as Substrate
Site PTM Type Enzyme
K97 Ubiquitination
S99 Phosphorylation P17612 (PRKACA)
K100 Ubiquitination
T107 Phosphorylation
K117 Ubiquitination
T125 Phosphorylation P27361 (MAPK3) , P28482 (MAPK1) , Q13627 (DYRK1A) , P06493 (CDK1)
S133 Phosphorylation
S144 Phosphorylation Q05513 (PRKCZ)
Y153 Phosphorylation P00519 (ABL1) , A0A173G4P4 (Abl fusion)
S175 Phosphorylation
S183 Phosphorylation P17612 (PRKACA)
K189 Ubiquitination
S195 Phosphorylation P17612 (PRKACA)
S196 Phosphorylation Q9Y243 (AKT3) , P31751 (AKT2) , P31749 (AKT1) , Q13237 (PRKG2)
K204 Ubiquitination
T208 Phosphorylation
K210 Ubiquitination
K211 Ubiquitination
Y251 Phosphorylation
K278 Ubiquitination
T301 Phosphorylation
S302 Phosphorylation
S307 Phosphorylation
S310 Phosphorylation
K394 Ubiquitination
Y397 Phosphorylation
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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