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

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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:

A synthesized peptide derived from human Caspase 9 (Cleaved-Asp330).

Uniprot:
Gene(ID):
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

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.

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=8.5]

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=4.8]

5). Lutein inhibits glutamate-induced apoptosis in HT22 cells via the Nrf2/HO-1 signaling pathway. Frontiers in neuroscience, 2024 (PubMed: 39193525) [IF=4.3]

6). 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=4.2]

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.

7). 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=3.5]

Application: WB    Species: human    Sample: A172 cells

FIGURE 8 Lentiviral knock-down of Ankfy1 expression in A172 cells promotes apoptosis.

8). Vascular endothelial-derived Von Willebrand factor inhibits lung cancer progression through the αvβ3/ERK1/2 axis. Toxicology and Applied Pharmacology, 2023 (PubMed: 37068611) [IF=3.3]

9). 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=3.3]

10). 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]

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Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

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