Cleaved-Caspase 9 (Asp330) Antibody - #AF5244

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Product: Cleaved-Caspase 9 (Asp330) Antibody
Catalog: AF5244
Description: Rabbit polyclonal antibody to Cleaved-Caspase 9 (Asp330)
Application: WB IHC IF/ICC
Reactivity: Human
Mol.Wt.: 10 kDa; 46kD(Calculated).
Uniprot: P55211
RRID: AB_2837730

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:1000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human
Clonality:
Polyclonal
Specificity:
Cleaved-Caspase 9 (Asp330) Antibody detects endogenous levels of fragment of activated Caspase 9 resulting from cleavage adjacent to Asp330.
RRID:
AB_2837730
Cite Format: Affinity Biosciences Cat# AF5244, RRID:AB_2837730.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
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.
Alias:

Fold/Unfold

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;

Immunogens

Immunogen:
Uniprot:

P55211(CASP9_HUMAN) >>Visit HPA database.

Gene(ID):
CASP9(842)
Expression:
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

PTMs - 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

Research Backgrounds

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.

PTMs:

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.

Family&Domains:

Belongs to the peptidase C14A family.

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 > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.

· Human Diseases > Neurodegenerative diseases > Parkinson's disease.

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Neurodegenerative diseases > Huntington's disease.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Colorectal cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Endometrial cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Small cell lung cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Non-small cell lung cancer.   (View pathway)

· Human Diseases > Cardiovascular diseases > Viral myocarditis.

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

References

1). CircGPR137B/miR-4739/FTO feedback loop suppresses tumorigenesis and metastasis of hepatocellular carcinoma. Molecular Cancer, 2022 (PubMed: 35858900) [IF=37.3]

Application: WB    Species: Human    Sample: HepG2 and Hep3B cells

Fig. 6 FTO is regulated by circGPR137B/miR-4739 axis in HCC. A Schematic representation of the binding sites between miR-4739 and FTO 3’UTR. B Comparison of the luciferase activity of WT or Mut FTO 3’UTR after treatment with miR-4739 mimics in HepG2 and Hep3B cells. C, D RT-qPCR and western blot analysis of the expression of FTO, p21, p27, cleaved caspase-3/− 9, N-cadherin, E-cadherin and vimentin after the transfection with circGPR137B lentiviruses and (or) miR-4739 mimics in HepG2 and Hep3B cells. E, F, G MTT, colony formation and transwell analysis of the cell proliferation, colony number and cell invasion after the transfection with FTO plasmids and (or) miR-4739 mimics in HepG2 and Hep3B cells. Data are the means ± SEM of three experiments. *P < 0.05, **P < 0.01, ***P < 0.001
2). Cytotoxicity of adducts formed between quercetin and methylglyoxal in PC-12 cells. Food Chemistry, 2021 (PubMed: 33706136) [IF=8.8]

Application: WB    Species: Rat    Sample: PC-12 cells

Fig. 5. Effect of treatments of MGO, Que-mono-MGO, and Que-di-MGO on the expression levels of apoptotic markers and components of AKT and Nrf2-HO-1/NQO-1 signaling pathways. Significant differences (p < 0.05) between samples of different treatments are marked with different letters on each column.
3). Photoprotective Effects of Dendrobium nobile Lindl. Polysaccharides against UVB-Induced Oxidative Stress and Apoptosis in HaCaT Cells. International Journal of Molecular Sciences, 2023 (PubMed: 37047098) [IF=5.6]

Application: IF/ICC    Species: Human    Sample: HaCaT cells

Figure 8 DNPs inhibit UVB-induced apoptosis-related proteins expression in HaCaT cells. (A) Representative fluorescence images showing cleaved caspase-9 staining and (B) cleaved caspase-3 staining. Scale bar = 100 μm (A)/20 μm (B); Cleaved caspase-9 and cleaved caspase-3 positive cells were counted (C,D). ### p < 0.001 vs. control;
4). Discovery of petroleum ether extract of eclipta targeting p53/Fas pathway for the treatment of chemotherapy-induced alopecia: Network pharmacology and experimental validation. Journal of ethnopharmacology, 2024 (PubMed: 38844249) [IF=5.4]
5). ADAM28 from both endothelium and gastric cancer cleaves von Willebrand Factor to eliminate von Willebrand Factor-induced apoptosis of gastric cancer cells. European Journal of Pharmacology, 2021 (PubMed: 33675784) [IF=5.0]

Application: WB    Species: Human    Sample: SGC7901 cells

Fig. 5. In the co-culture system, ADAM28 derived from endothelium reduces the apoptosis of lower ventricular gastric cancer cells. (A) Western blot analysis of ADAM28 expression in HUVECs with ADAM28 overexpression or knockdown (***P < 0.001). (B–C) Flow cytometry was used to analyze the effects of HUVEC, HUVEC-ADAM28-KD and HUVEC-ADAM28-OE on the apoptosis of gastric cancer cells in the lower ventricle. Percentage of apoptotic cells compared to control was quantitated by mean fluorescence intensity and was shown in right graphs (***P < 0.001). (D) When HUVEC, HUVEC-ADAM28-KD and HUVEC-ADAM28-OE were added into the upper chamber, Western blot analysis was performed on SGC7901 cells in the lower chamber with antibodies against Casp3/c-Casp3 and Casp9/cCasp9. β-actin was used as a loading control. Each bar represented the mean ± S.D. of three independent experiments.
6). Purkinje Cell Degeneration and Motor Coordination Deficits in a New Mouse Model of Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay. Frontiers in Molecular Neuroscience, 2017 (PubMed: 28588446) [IF=4.8]

Application: WB    Species: human    Sample: A172 cells

FIGURE 8 Lentiviral knock-down of Ankfy1 expression in A172 cells promotes apoptosis.
7). Vascular endothelial-derived Von Willebrand factor inhibits lung cancer progression through the αvβ3/ERK1/2 axis. Toxicology and Applied Pharmacology, 2023 (PubMed: 37068611) [IF=3.8]
8). Involvement of estrogen receptor activation in kaempferol-3-O-glucoside's protection against aging-related cognition impairment and microglial inflammation. Experimental cell research, 2023 (PubMed: 37926343) [IF=3.7]
9). Inhibition of P21-activated kinase 1 promotes vascular smooth muscle cells apoptosis through reduction of phosphorylation of Bad. American Journal of Hypertension, 2023 (PubMed: 36634025) [IF=3.2]
10). Lutein inhibits glutamate-induced apoptosis in HT22 cells via the Nrf2/HO-1 signaling pathway. Frontiers in neuroscience, 2024 (PubMed: 39193525) [IF=3.2]

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