Cleaved-Caspase 9 (Asp353) Antibody - #AF5240

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产品: Cleaved-Caspase 9 (Asp353) 抗体
货号: AF5240
描述: Rabbit polyclonal antibody to Cleaved-Caspase 9 (Asp353)
应用: WB IHC IF/ICC
反应: Mouse, Rat
分子量: 17/38 kDa(mature), 47kDa(precursor); 46kD(Calculated).
蛋白号: P55211
RRID: AB_2837726

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实验方案
WB Handbook
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产品描述

来源:
Rabbit
应用:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: 适用于变性蛋白样本的免疫印迹检测. IHC: 适用于组织样本的石蜡(IHC-p)或冰冻(IHC-f)切片样本的免疫组化/荧光检测. IF/ICC: 适用于细胞样本的荧光检测. ELISA(peptide): 适用于抗原肽的ELISA检测.

反应:
Mouse,Rat
克隆:
Polyclonal
特异性:
Cleaved-Caspase 9 (Asp353) Antibody detects endogenous levels of fragment of activated Caspase 9 resulting from cleavage adjacent to Asp353.
RRID:
AB_2837726
引用格式: Affinity Biosciences Cat# AF5240, RRID:AB_2837726.
偶联:
Unconjugated.
纯化:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
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.
别名:

展开/折叠

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;

抗原和靶标

免疫原:
Uniprot:

P55211(CASP9_HUMAN) >>Visit HPA database.

基因/基因ID:
CASP9(842)
表达:
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.

描述:
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).
序列:
MDEADRRLLRRCRLRLVEELQVDQLWDALLSRELFRPHMIEDIQRAGSGSRRDQARQLIIDLETRGSQALPLFISCLEDTGQDMLASFLRTNRQAAKLSKPTLENLTPVVLRPEIRKPEVLRPETPRPVDIGSGGFGDVGALESLRGNADLAYILSMEPCGHCLIINNVNFCRESGLRTRTGSNIDCEKLRRRFSSLHFMVEVKGDLTAKKMVLALLELAQQDHGALDCCVVVILSHGCQASHLQFPGAVYGTDGCPVSVEKIVNIFNGTSCPSLGGKPKLFFIQACGGEQKDHGFEVASTSPEDESPGSNPEPDATPFQEGLRTFDQLDAISSLPTPSDIFVSYSTFPGFVSWRDPKSGSWYVETLDDIFEQWAHSEDLQSLLLRVANAVSVKGIYKQMPGCFNFLRKKLFFKTS

翻译修饰 - P55211 作为底物

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

研究背景

功能:

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.

翻译修饰:

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.

组织特异性:

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.

亚基结构:

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.

蛋白家族:

Belongs to the peptidase C14A family.

研究领域

· 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)

文献引用

1). Upregulation of BCL-2 by acridone derivative through gene promoter i-motif for alleviating liver damage of NAFLD/NASH. NUCLEIC ACIDS RESEARCH (PubMed: 32710621) [IF=14.9]

Application: WB    Species: human    Sample: HepG2

Figure 4. Effect of A22 on anti-apoptosis in 0.5 mM palmitic acid oil (PA) induced cell model. (A) Effect of A22 on cell viability for anti-apoptotic protective effect. (B) Effect of A22 on transcription of BCL-2 and BAX with measurement of mRNA levels. (C) Effect of A22 on protein expressions related with apoptosis (left), which were quantitatively analyzed (right). All the experiments were repeated for three times.
2). Formation of di-cysteine acrolein adduct decreases cytotoxicity of acrolein by ROS alleviation and apoptosis intervention. Journal of Hazardous Materials (PubMed: 31780296) [IF=13.6]

Application: WB    Species: Human    Sample: HBE (A) and Caco-2 (B) cells

Fig. 8 Effect of acrolein (ACR) or its adduct ACR-di-Cys with/without addition of 1mM cysteine on the cleavage of caspase-3, caspase-8 and PARP in HBE (A) and Caco-2 (B) cells. Left: western blot. Right: The data were expressed as relative intensity to GAPDH; Error bars represent standard deviation (n=3); Different letters indicate significant differences (p < 0.05) between treatments. 42 J

Application: WB    Species: Human    Sample: HBE and Caco-2 cells

Fig. 9 Effect of acrolein (ACR) or its adduct ACR-di-Cys with/without addition of 1mM cysteine on the expression of Bax, Bcl-2, cytochrome c and the cleavage of caspase-9 in HBE (A) and Caco-2 (B) cells. Left: western blot. Right: The data were expressed as relative intensity to GAPDH; Error bars represent standard deviation (n=3); Different letters indicate significant differences (p < 0.05) between treatments.
3). The role of the apoptosis-related protein BCL-B in the regulation of mitophagy in hepatic stellate cells during the regression of liver fibrosis. EXPERIMENTAL AND MOLECULAR MEDICINE (PubMed: 30635551) [IF=12.8]

Application: WB    Species: mouse    Sample: HSC apoptosis

Fig. 3| Activation of mitophagy induces apoptosis in HSCs. a Mitochondrial DNA (mtDNA) measured by PicoGreen staining in LX2 cells; scale bar,10 μm. b Representative western blots of TOM20 with GAPDH serving as the internal reference. Bar graph represents the mean ± SEM. *P < 0.05 vs.the indicated groups. c TUNEL staining of LX2 cells from the indicated groups; scale bar, 25 μm. Bar graph represents the mean ± SEM. *P < 0.05 vs.the indicated groups. D Annexin V-FITC/PI double-staining and flow cytometry analysis of LX2 cells. Bar graph represents the mean ± SEM. **P < 0.01 vs. the indicated groups. e Representative western blots of cleaved caspase3, cleaved caspase9, collagen I and α-SMA. Bar graph represents the mean± SEM of three different experiments. *P < 0.05 and **P < 0.01 vs. the indicated groups
4). Cell-cycle arrest and mitochondria-dependent apoptosis induction in T-47D cells by the capsular polysaccharide from the marine bacterium Kangiella japonica KMM 3897. Carbohydrate Polymers (PubMed: 37659798) [IF=11.2]
5). Bacillus spore-based oral carriers loading curcumin for the therapy of colon cancer. JOURNAL OF CONTROLLED RELEASE (PubMed: 29274436) [IF=10.8]

Application: WB    Species: human    Sample: HT-29 cells

Figure.5 | Apoptosis detection of HT-29 colon cancer cells. (A) Apoptosis detection of HT-29 cells in different groups by flow cytometry, SFM without drug as control; (B) Apoptosis rates of HT-29 cells in different groups. (mean value ± SD, n=3, **p < 0.01, ***p < 0.001, compared with the control group); (C) Western blotting of the Bcl-2, p53, cleaved caspase-9, cleaved caspase-8,cleaved caspase-3; (D) Relative amount of these apoptosis-related proteins in different groups(mean value ± SD, n=3, *p < 0.05, **p < 0.01, ***p < 0.001, compared with the control group).
6). Rescuing ischemic stroke by biomimetic nanovesicles through accelerated thrombolysis and sequential ischemia-reperfusion protection. Acta Biomaterialia (PubMed: 34902617) [IF=9.7]
7). Morinda officinalis oligosaccharides mitigate depression-like behaviors in hypertension rats by regulating Mfn2-mediated mitophagy. Journal of Neuroinflammation (PubMed: 36765376) [IF=9.3]

Application: WB    Species: Rat    Sample:

Fig. 5 Autophagy inhibitors aggravated neuronal toxicity inhibited by Morinda officinalis oligosaccharides by preventing mitochondrial damage. A Tumor necrosis factor α, interleukin (IL)-6, and IL-1β levels in astrocyte supernatants were determined by enzyme-linked immunosorbent assay. Intracellular (B) and mitochondrial (C) reactive oxygen species levels were assessed using the DCFDA probe and MitoSOX probe. D Mitochondrial membrane potential was assessed using the JC-1 probe. E Isolation and identification of primary neurons by light microscopy and fluorescent staining for neuron-specific enolase. F Cell viability of neurons was determined by cell counting kit-8 assay. G Apoptosis of neurons was determined by flow cytometry. H Apoptosis-related protein levels (cleaved caspase-3 and cleaved caspase-9) in neurons were determined by western blotting. Data are shown as the mean ± standard deviation of three independent experiments. **p 
8). Iron induces B cell pyroptosis through Tom20–Bax–caspase–gasdermin E signaling to promote inflammation post-spinal cord injury. Journal of Neuroinflammation (PubMed: 37480037) [IF=9.3]

Application: WB    Species: Mouse    Sample: B cells

Fig. 3 Pyroptosis induced by iron accumulation may occur in splenic B cells after SCI. a Three days after SCI, serum samples were collected for quantification of the protein expression levels of MCP-1, IL-1β, IL-6, and TNF-α by ELISA. b Three days after SCI, injured spinal cord homogenates were collected for quantification of the protein expression levels of MCP-1, IL-1β, IL-6, and TNF-α by ELISA. c Three days after SCI, the spleen was stained with Prussian blue to detect the level of iron ion. d Three days after SCI, serum samples were collected to quantify the concentration of iron ions. e Three days after SCI, B cell lysates were collected to quantify the concentration of iron ions. f Detection of Tom20-Bax-caspase-GSDME pathway-related protein expression in B cells by western blot. g The knock-down efficiency of AAV on Tom20 in B cells was verified. h, l, m The expression levels of MCP-1, IL-1 β, IL-6, TNF- α, IgG and IgM in serum of different groups were detected by ELISA. i–k Three days after SCI, the spleens of mice were observed, and the spleen length and organ index of different groups were compared. n, o, q The spleen was taken for HE staining, Prussian blue staining, and immunofluorescence. CD19+ was red, dapi was blue, and merge was combined. p Detection of nerve evoked potentials in different groups of mice. r Detection of the expression of Tom20-Bax-caspsae-GSDME pathway-related proteins in different groups of B cells. All data are expressed as the mean ± SD (n ≥ 3 replicates per group). ns P > 0.05, * P 
9). LNC473 regulating APAF1 IRES-dependent translation via competitive sponging miR574 and miR15b: Implications in colorectal cancer. Molecular Therapy-Nucleic Acids (PubMed: 32784109) [IF=8.8]

Application: WB    Species: Human    Sample: HCT116 and SW480 cells

Figure 6. LNC473-miR574/miR15b-APAF1 Signaling Axis in HCT116 and SW480 Cells (A) IF and ISH combination assay revealing the co-localization of APAF1 protein with included ncRNAs in HCT116 cells. Scale bars, 5 mm. (B and C) The levels of APAF1 mRNA (B) and protein (C) were determined after interfering LNC473 expression in HCT116 and SW480 cells by qPCR and IF assays. Scale bars, 20 mm. (D) The expression of apoptosis-related proteins including APAF1 was detected after interfering LNC473 expression in HCT116 and SW480 cells by western blot assay. (E and F) Rescue experiments showing the APAF1 levels in HCT116 and SW480 cells with exposure to the co-transfection of pcDH-LNC473 vector and miR574-5p or miR15b-5p mimic by qPCR (E) and western blot (F) assays. (G) Pattern diagram of APAF1-CDS and APAF1-IRES-CDS vectors. (H and I) APAF1 protein expression was determined in HCT116 and SW480 cells treated with APAF1-IRES-CDS vector, or pcDH-LNC473 and APAF1-IRES-CD co-transfection by western blot (H) and IF assays (I). Scale bars, 5 mm. (J) The percentage (%) of cell apoptosis in cells upon co-overexpressing APAF1-CDS or APAF1-IRE-CDS and LNC473 as assayed by flow cytometry. All tests were performed at least three times. Data were expressed as mean ± SD. ns (nonsignificant), p > 0.05; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
10). Long noncoding RNAs regulated spermatogenesis in varicocele‐induced spermatogenic dysfunction. CELL PROLIFERATION (PubMed: 35297519) [IF=8.5]

Application: WB    Species: rat    Sample: Testicular

FIGURE 13| Validation of regulated signalling pathways, spermatogenic cell apoptosis and proliferation, and meiotic spermatocytes by Western blot. Representative Western blot images of PI3K, Akt, p-Akt, caspase-9, Bcl-2, Bax, PCNA, PLZF, REC8, STRA8, and SYCP3.
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