Cleaved-Caspase 3 (Asp175), p17 Antibody - #AF7022

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产品: Cleaved-Caspase 3 (Asp175), p17 抗体
货号: AF7022
描述: Rabbit polyclonal antibody to Cleaved-Caspase 3 (Asp175), p17
应用: WB IHC IF/ICC
反应: Human, Mouse, Rat, Bovine
预测: Pig, Zebrafish, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus
分子量: 17kDa; 32kD(Calculated).
蛋白号: P42574
RRID: AB_2835326

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WB Handbook
IHC Protocol
IF/ICC Protocol

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:
Human,Mouse,Rat,Bovine
预测:
Pig(100%), Zebrafish(0%), Horse(86%), Sheep(100%), Rabbit(86%), Dog(100%), Xenopus(86%)
克隆:
Polyclonal
特异性:
Cleaved-Caspase 3 (Asp175,p17) Antibody detects endogenous levels of fragment of activated Caspase 3 resulting from cleavage adjacent to Asp175.
RRID:
AB_2835326
引用格式: Affinity Biosciences Cat# AF7022, RRID:AB_2835326.
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.Stable for 12 months from date of receipt. Store at -20 °C. Stable for 12 months from date of receipt.
别名:

展开/折叠

A830040C14Rik; Apopain; CASP-3; CASP3; CASP3_HUMAN; Casp3a; Caspase 3; Caspase 3, apoptosis-related cysteine peptidase; Caspase 3, apoptosis-related cysteine protease; Caspase 3, apoptosis-related cysteine protease a; Caspase-3 subunit p12; CC3; CPP-32; CPP32; CPP32B; Cysteine protease CPP32; EC 3.4.22.56; LICE; mldy; OTTHUMP00000165052; OTTHUMP00000165053; OTTHUMP00000165054; PARP cleavage protease; Procaspase3; protein Yama; SCA 1; SCA-1; SREBP cleavage activity 1; Yama;

抗原和靶标

免疫原:
Uniprot:

P42574(CASP3_HUMAN) >>Visit HPA database.

基因/基因ID:
CASP3(836)
表达:
P42574 CASP3_HUMAN:

Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

描述:
This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce 2 subunits, large and small, that dimerize to form the active enzyme.
序列:
MENTENSVDSKSIKNLEPKIIHGSESMDSGISLDNSYKMDYPEMGLCIIINNKNFHKSTGMTSRSGTDVDAANLRETFRNLKYEVRNKNDLTREEIVELMRDVSKEDHSKRSSFVCVLLSHGEEGIIFGTNGPVDLKKITNFFRGDRCRSLTGKPKLFIIQACRGTELDCGIETDSGVDDDMACHKIPVEADFLYAYSTAPGYYSWRNSKDGSWFIQSLCAMLKQYADKLEFMHILTRVNRKVATEFESFSFDATFHAKKQIPCIVSMLTKELYFYH

种属预测

种属预测:

score>80的预测可信度较高，可尝试用于WB检测。*预测模型主要基于免疫原序列比对，结果仅作参考，不作为质保凭据。

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Horse
86
Xenopus
86
Rabbit
86
Chicken
71
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

翻译修饰 - P42574 作为底物

Site PTM Type Enzyme
M1 Acetylation
T4 Phosphorylation
S7 Phosphorylation
S10 Phosphorylation
K11 Acetylation
K11 Ubiquitination
S12 Phosphorylation
K14 Ubiquitination
K19 Ubiquitination
S24 Phosphorylation
S26 Phosphorylation
S29 Phosphorylation
Y41 Phosphorylation
K57 Ubiquitination
S65 Phosphorylation
T67 Phosphorylation
K82 Acetylation
K82 Ubiquitination
K88 Ubiquitination
K105 Ubiquitination
K138 Ubiquitination
S150 Phosphorylation Q16539 (MAPK14)
T152 Phosphorylation
C163 S-Nitrosylation
T174 Phosphorylation
S176 Phosphorylation
K210 Ubiquitination
K229 Ubiquitination
S249 Phosphorylation
K260 Ubiquitination
T270 Phosphorylation

研究背景

功能:

Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-|-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. Triggers cell adhesion in sympathetic neurons through RET cleavage.

翻译修饰:

Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.

S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.

细胞定位:

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
组织特异性:

Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.

亚基结构:

Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 17 kDa (p17) and a 12 kDa (p12) subunit. Interacts with BIRC6/bruce.

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 > MAPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > TNF signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).

· 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 > Epithelial cell signaling in Helicobacter pylori infection.

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.

· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.

· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.

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

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

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

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

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

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

· Human Diseases > Cardiovascular diseases > Viral myocarditis.

· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity.   (View pathway)

· Organismal Systems > Immune system > IL-17 signaling pathway.   (View pathway)

· Organismal Systems > Nervous system > Serotonergic synapse.

文献引用

1). CircGPR137B/miR-4739/FTO feedback loop suppresses tumorigenesis and metastasis of hepatocellular carcinoma. Molecular Cancer (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). 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.
3). Antitumor activity of aumolertinib, a third-generation EGFR tyrosine kinase inhibitor, in non-small-cell lung cancer harboring uncommon EGFR mutations. Acta Pharmaceutica Sinica B (PubMed: 37425047) [IF=14.5]

Application: WB    Species: Mouse    Sample: tumor tissue

Figure 6 Antitumor activity of aumolertinib in the PDX model harboring EGFR H773-V774insNPH mutation. Tumor volume (A), tumor weight (B), average body weights of mice at treatment start (Day 0) and study end (Day 26) (C), and the photograph of tumors (D) in a PDX LU0387 (EGFR H773_N774insNPH) NSCLC tumor model in mice over 26-day treatment with aumolertinib. The vehicle was used as the control. (E, F) Expression of pEGFR, EGFR, pAKT, AKT, pERK, ERK, PCNA, cleaved PARP, PARP, cleaved caspase-3, and caspase-3 in tumor tissue after administration of EGFR-TKIs. Tumors were obtained and lysed at 6 h after the last treatment. Data are presented as the ratio relative to the vehicle group. The data of (A), (B), (C), and (F) are presented as the mean ± SEM (n = 6/group) and compared with the vehicle (∗P < 0.05; ∗∗P < 0.01). PDX, patient-derived xenografts; EGFR, epidermal growth factor receptor; TKIs, tyrosine kinase inhibitors; NPH, H773-V774insNPH.
4). Cyanidin-3-O-glucoside restores spermatogenic dysfunction in cadmium-exposed pubertal mice via histone ubiquitination and mitigating oxidative damage. JOURNAL OF HAZARDOUS MATERIALS (PubMed: 31796358) [IF=13.6]

Application: WB    Species: Mice    Sample: testis

Figure 9 The representative photographs and grayscale analysis of proteins in testis from mice treated for 30 days involved in MAPK signaling pathway. (A) p53 and p-p53. (B) Bax, Bcl-2, and Bad. (C) Total Caspase 3 and Cleaved-Caspase 3. (D) Full PARP and Cleaved-PARP. Comparison between all groups was evaluated through One-way ANOVA. Mean±SD. n=4. **p<0.01, *p<0.05, compared with control group. ##p<0.01, #p<0.05, compared with Cd group. ns, not significant.
5). Prenatal exposure to environmentally relevant levels of PBDE-99 leads to testicular dysgenesis with steroidogenesis disorders. JOURNAL OF HAZARDOUS MATERIALS (PubMed: 34879533) [IF=13.6]
6). 2,3′,4,4′,5-Pentachlorobiphenyl induces mitochondria-dependent apoptosis mediated by AhR/Cyp1a1 in mouse germ cells. Journal of Hazardous Materials (PubMed: 37055962) [IF=13.6]
7). HSP70 attenuates compression-induced apoptosis of nucleus pulposus cells by suppressing mitochondrial fission via upregulating the expression of SIRT3. EXPERIMENTAL AND MOLECULAR MEDICINE (PubMed: 35338257) [IF=12.8]

Application: WB    Species: Rat    Sample: NP cells

Fig. 4 HSP70 inhibited the mitochondrial apoptotic pathway. a, b The MMP of NP cells measured by flow cytometry using JC-1 staining. c, d The production of mtROS in NP cells measured by flow cytometry using MitoSOX Red staining. e, f The production of cellular ROS in NP cells measured by flow cytometry using DCFH-DA staining. g The effects of TRC on ATP production in NP cells. h The MMP and production of mtROS and cellular ROS in NP cells measured by flow cytometry. i The effects of VER on ATP production in NP cells treated with TRC. j The effects of TRC on the expression of cleaved caspase 3, cleaved caspase 9, Bax, and Bcl-2. k The effects of VER on the expression of cleaved caspase 3, cleaved caspase 9, Bax, and Bcl-2 in NP cells treated with TRC. (N = 3, *P < 0.05; **P < 0.01; ***P < 0.001).

Application: IHC    Species: Rat    Sample: NP cells

Fig. 4 HSP70 inhibited the mitochondrial apoptotic pathway. a, b The MMP of NP cells measured by flow cytometry using JC-1 staining. c, d The production of mtROS in NP cells measured by flow cytometry using MitoSOX Red staining. e, f The production of cellular ROS in NP cells measured by flow cytometry using DCFH-DA staining. g The effects of TRC on ATP production in NP cells. h The MMP and production of mtROS and cellular ROS in NP cells measured by flow cytometry. i The effects of VER on ATP production in NP cells treated with TRC. j The effects of TRC on the expression of cleaved caspase 3, cleaved caspase 9, Bax, and Bcl-2. k The effects of VER on the expression of cleaved caspase 3, cleaved caspase 9, Bax, and Bcl-2 in NP cells treated with TRC. (N = 3, *P < 0.05; **P < 0.01; ***P < 0.001).
8). 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
9). PTBP1 knockdown promotes neural differentiation of glioblastoma cells through UNC5B receptor. Theranostics (PubMed: 35664063) [IF=12.4]

Application: WB    Species: Human    Sample: U251 cells

Figure 7 A DAPK1 inhibitor prevents the PTBP1 knockdown induced reprogramming. (A-E) Western blot study of NTN1, DAPK1, P-DAPK1(Ser308), P-P53(ser20), and caspase 3(p17) protein in U251 cells infected with sh-Luci and sh-PTBP1 for 3 and 7 days. (n = 3). As an internal reference protein, GAPDH was used. (F) Activity of PP2A during reprogramming. (n = 3). (G-I) TUJ1 and KI67 positive rates of six distinct groups constituted of lentivirus (sh-Luci or sh-PTBP1) and TC-DAPK6 (vehicle, 100 nM, 250 nM, and 500 nM; nine random fields from triplicate samples were recorded for quantification; TUJ1+ (%) = TUJ1+ M-cherry+/M-cherry+; KI67+ (%) = KI67+ M-cherry+/M-cherry+; M-cherry+ cells = 156-553 for each condition). The data are presented as mean ± SD. *P < 0.05, *** P < 0.001 vs. sh-Luci-3d or sh-PTBP1 + vehicle group. Dpi (d): days post infection; ND: not detected; NS: no significance. Scale: 100 µm.
10). TFE3, a potential therapeutic target for Spinal Cord Injury via augmenting autophagy flux and alleviating ER stress. Theranostics (PubMed: 32802192) [IF=12.4]

Application: WB    Species: mice    Sample: spinal cord

Figure 2. Inhibition of autophagy activity aggravates ER stress-induced apoptosis after SCI. (A) Western blot analysis of autophagy flux markers (ATG5, Beclin1, VPS34, C-CTSD, SQSTM1/p62, UB and LC3) and ER stress-induced apoptosis markers (CHOP, CASP12, C-CASP12, C-CASP3) in spinal cord lesions from ATG5-/+ mice and ATG5+/+ mice with and without SCI, at Day3 after SCI. (B, C) The levels of the autophagy flux makers from (A) normalized to loading control GAPDH. (D) The expressions of ER stress-induced apoptosis markers from (A) normalized to loading control GAPDH. (E) Image (30×) of spinal cord sections from the indicated groups at Day 3 stained with antibodies against LC3II/NeuN, p62/NeuN, CHOP/NeuN,and CASP3/NeuN, respectively. Scale bar: 25 µm. (F) Quantification of immunofluorescence data from (E) showing the mean number of LC3II in motor neurons at the spinal cord. (G-I) Quantification of immunofluorescence data from (E) showing the mean optical density of p62, CHOP, and CASP3, respectively, in motor neurons of spinal cord. n=6, ns stands for not significant, *P<0.05, **P<0.01.
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