Cleaved-Caspase 3 (Asp175), p17 Antibody - #BF0711

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製品: Cleaved-Caspase 3 (Asp175), p17 Antibody
カタログ: BF0711
タンパク質の説明: Mouse monoclonal antibody to Cleaved-Caspase 3 (Asp175), p17
アプリケーション: WB IHC IF/ICC
反応性: Human, Mouse, Rat
分子量: 17kDa; 32kD(Calculated).
ユニプロット: P42574
RRID: AB_2846190

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製品説明

ソース:
Mouse
アプリケーション:
WB(1:500-1:2000), IHC(1:200-1:1000), IF(1:50-1:200)
*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.

反応性:
Human,Mouse,Rat
クローナリティ:
Monoclonal [AFB17776]
特異性:
Caspase 3 (p17,Cleaved-Asp175) Antibody detects endogenous levels of fragment of activated Caspase 3 resulting from cleavage adjacent to Asp175.
RRID:
AB_2846190
引用形式: Affinity Biosciences Cat# BF0711, RRID:AB_2846190.
コンジュゲート:
Unconjugated.
精製:
Affinity-chromatography.
保存:
Mouse IgG1 in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

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;

免疫原

免疫原:

Purified recombinant fragment of human Caspase 3 expressed in E. Coli.

Uniprot:

P42574(CASP3_HUMAN) >>Visit HPA database.

遺伝子(ID):
CASP3(836)
発現特異性:
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.

タンパク質配列:
MENTENSVDSKSIKNLEPKIIHGSESMDSGISLDNSYKMDYPEMGLCIIINNKNFHKSTGMTSRSGTDVDAANLRETFRNLKYEVRNKNDLTREEIVELMRDVSKEDHSKRSSFVCVLLSHGEEGIIFGTNGPVDLKKITNFFRGDRCRSLTGKPKLFIIQACRGTELDCGIETDSGVDDDMACHKIPVEADFLYAYSTAPGYYSWRNSKDGSWFIQSLCAMLKQYADKLEFMHILTRVNRKVATEFESFSFDATFHAKKQIPCIVSMLTKELYFYH

PTMs - P42574 基板として

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

機能:

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.

PTMs:

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.

細胞の位置付け:

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

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.

サブユニット構造:

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.

タンパク質ファミリー:

Belongs to the peptidase C14A family.

研究領域

· 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.

参考文献

1). Π electron-stabilized polymeric micelles potentiate docetaxel therapy in advanced-stage gastrointestinal cancer. BIOMATERIALS, 2021 (PubMed: 33069116) [IF=14.0]

Application: IF/ICC    Species: Mice    Sample: Tumor

Figure 5 DTX-PM improve DTX efficacy in a PDX model of GI cancer. A: Tumor growth inhibition upon treatment with PBS, and with 10 mg/kg of free DTX and DTX-PM. B: Tumor weights at the end of the experiment, showing significantly more efficient tumor inhibition for DTX-PM than for free DTX. C-E: Immunofluorescence analysis of cellular proliferation (Ki67) and apoptosis induction (cleaved caspase 3), confirming that DTX-PM was more effective than free DTX. F and G: Individual PDX tumor growth curves in the mice treated with DTX and DTX-PM as compared to PBS controls, exemplifying the superior efficacy of DTX-PM. H and I: PDX tumors were analyzed by means of immunofluorescence microscopy, showing a substantially increased ratio of M1-like to M2-like TAM upon DTX-PM treatment.
2). Targeting NUPR1-dependent stress granules formation to induce synthetic lethality in KrasG12D-driven tumors. EMBO molecular medicine, 2024 (PubMed: 38360999) [IF=11.1]

Application: IF/ICC    Species: Mouse    Sample:

Figure 7 NUPR1 inhibition induced cell death by apoptosis in KrasG12D expressing cells in vivo. Immunohistofluorescence staining was performed on histologic sections of the pancreas of the different experimental groups. Rabbit anti-KrasG12D primary antibody and mouse anti-cleaved caspase 3 antibody was used. DNA was stained with DAPI. Pearson coefficient between both channels was calculated by JACoP, ImageJ. Data represent mean ± SD, ANOVA with Sidak correction (n = 5 mice).
3). Oral P. gingivalis impairs gut permeability and mediates immune responses associated with neurodegeneration in LRRK2 R1441G mice. Journal of Neuroinflammation, 2020 (PubMed: 33213462) [IF=9.3]

Application: WB    Species: mice    Sample: R1441G cells and FVBN cells

Fig.1 c Representative images of western blots of cleaved caspase-3 protein levels and quantitative analysis, which were done with the SN obtained from F + Pg and 1441 + Pg mice compared to F + C and 1441 + C mice. n = 4, **P < 0.01. Two-way ANOVA and Tukey’s test were used for analysis.
4). Smooth muscle-specific HuR knockout induces defective autophagy and atherosclerosis. Cell Death & Disease, 2021 (PubMed: 33837179) [IF=9.0]

Application: WB    Species: mouse    Sample: SMCs

Fig. 3| Loss of HuR promoted apoptosis in atherosclerosis. A Immunofluorescent staining of aortic roots from CTR and HuRSMKO mice to determine TUNEL-positive VSMCs. Red puncta denotes TUNEL-positive cells. Green region denotes α-SMA. Scale bar = 20 μm. B Immunohistochemical staining of cleaved caspase-3 in aortic roots (n = 6). Scale bar = 50 μm. C Serum lipid profiles (total cholesterol [TC], triglycerides [TG], high-density lipoprotein cholesterol [HDL-C], and low-density lipoprotein cholesterol [LDL-C]) (n = 6). D Western blot analysis of cleaved caspase-3 in control and HuR-deficient SMCs (n = 5).
5). Focal ischemic stroke modifies microglia-derived exosomal miRNAs: potential role of mir-212-5p in neuronal protection and functional recovery. Biological Research, 2023 (PubMed: 37789455) [IF=6.7]

Application: IF/ICC    Species: Rat    Sample: PC12 cells

Fig. 9 MiR-212-5p reduces neuronal apoptosis and attenuates axonal degeneration in PC12 cells following OGD/R. A Cleaved-Caspase 3 immunofluorescence staining (in green). Nuclei were stained with DAPI and are visualised in blue. Scale bar = 50 μm. B, C Nogo-A and NgR gene expression levels in PC12 cells. Gene expression levels were examined using qRT–PCR and normalised to β-actin. D-F Immunofluorescence staining was conducted to reveal the presence of Nogo-A, NgR and β III tubulin in PC12 cells. Nuclei were stained with DAPI and are visualised in blue. Scale bar = 50 μm. The data are presented as the means ± SEM (n = 5 per group). *P 
6). PLXNA2 knockdown promotes M2 microglia polarization through mTOR/STAT3 signaling to improve functional recovery in rats after cerebral ischemia/reperfusion injury. EXPERIMENTAL NEUROLOGY, 2021 (PubMed: 34474008) [IF=5.3]

Application: IF/ICC    Species: Mice    Sample: BV2 microglia cells

Fig. 8. The effects of PLXNA2 knockdown on the mTOR/STAT3 pathway in oxygen and glucose deprivation/reoxygenation (OGD/R)-stimulated BV2 microglia cells. (A) Immunofluorescence staining for PLXNA2 in OGD/R-stimulated BV2 microglia cells. Scale bar = 50 μm. (B) Immunofluorescence staining for cleaved caspase 3 in OGD/R-stimulated BV2 microglia cells. Scale bar = 50 μm. (C) Representative Western blot bands show the protein expression of PLXNA2, p-mTOR, mTOR, p-STAT3 and STAT3 in BV2 microglia cells. β-actin was used as internal control. (D–F) Quantification of immunoblots of PLXNA2, p-mTOR and p-STAT3. The data are presented as the mean ± SEM for each group (n = 4–6 per group). *p < 0.05, **p < 0.01 versus the sham + scrambled group, #p < 0.05, ##p < 0.01 versus the MCAO/ R + scrambled group.
7). Fibroblast growth factor 2 contributes to the effect of salidroside on dendritic and synaptic plasticity after cerebral ischemia/reperfusion injury. Aging (Albany NY), 2020 (PubMed: 32518214) [IF=5.2]

Application: IF/ICC    Species: rat    Sample:

Figure 8.| Sal attenuates neuronal apoptosis after MCAO/R. (A) Immunofluorescence staining of c-caspase 3 in sections from the ischemic penumbra in each group on day 7 post-MCAO/R (the scale bar is 20 μm).

Application: WB    Species: rat    Sample:

Figure 8.| Sal attenuates neuronal apoptosis after MCAO/R. (A) Immunofluorescence staining of c-caspase 3 in sections from the ischemic penumbra in each group on day 7 post-MCAO/R (the scale bar is 20 μm).. (B–E) Protein expression of c-caspase 3, Bcl-2 and Bax from the ischemic penumbra. (F–H) QPCR results of c-caspase 3, Bcl-2 and Bax expression. Values are expressed as the mean ± SD. #p < 0.05, ##p <0.01 vs. sham; *p < 0.05, **p < 0.01 vs. MCAO/R.
8). Tannic Acid Alleviates Lipopolysaccharide-Induced H9C2 Cell Apoptosis by Suppressing ROS-Mediated ER Stress. Molecular Medicine Reports, 2021 (PubMed: 34080663) [IF=3.4]

Application: WB    Species: rat    Sample: H9C2 cells

Figure 2.| TA alleviates the LPS‑induced apoptosis of H9C2 cells. (C) Protein expression levels of Bax, Bcl‑2, cleaved caspase‑9, cleaved caspase‑3 and β‑actin in H9C2 cells treated with LPS, TA or TA + LPS were determined by western blot analysis.

Application: WB    Species: Rat    Sample: H9C2 cells

Figure 6. TA attenuates LPS-induced apoptosis via suppressing ROS-mediated endoplasmic reticulum stress in H9C2 cells. (A) Whole cell lysates were prepared following stimulation with LPS, NAC and TA. The protein expression levels of apoptosis-associated proteins (p-JNK, JNK, Bax, cleaved caspase-9 and cleaved caspase-3) and an anti-apoptotic protein (Bcl-2) were detected in H9C2 cells treated with LPS, NAC + LPS, TA + LPS and TA + NAC + LPS by western blot analysis. Semi-quantification of the protein expression levels of (B) p-JNK/JNK and cleaved caspase-3, and (C) Bax/Bcl-2 and cleaved caspase-9 in H9C2 cells. The mRNA expression levels of (D) Bax, (E) caspase-3, (F) caspase-12 and (G) Cyt c in H9C2 cells normalized to Gapdh. Data are presented as the mean ± standard error of the mean (n=3). *P<0.05, **P<0.01 and ***P<0.001 vs. control; #P<0.05 and ##P<0.01 vs. LPS group; §P<0.05 and §§P<0.01 vs. TA + LPS. TA, tannic acid; LPS, lipopolysaccharide; NAC, N-acetylcysteine; p, phosphorylated; JNK, c-Jun N-terminal kinase; Cyt c, cytochrome c.

Application: WB    Species: Rat    Sample: H9C2 cells

Figure 2. TA alleviates the LPS-induced apoptosis of H9C2 cells. (A) Flow cytometry was used to determine cell apoptotic rate using Annexin V-FITC/PI staining. Apoptotic cells included Annexin V+/PI- and Annexin V+/PI+ cells. (B) Quantitative analysis of the apoptotic rate of cells treated with LPS, TA and TA + LPS. (C) Protein expression levels of Bax, Bcl-2, cleaved caspase-9, cleaved caspase-3 and β-actin in H9C2 cells treated with LPS, TA or TA + LPS were determined by western blot analysis. (D) Semi-quantification of the protein expression levels of Bax/Bcl-2, cleaved caspase-9 and cleaved caspase-3 in the LPS, TA and TA + LPS groups. Data are presented as the mean ± standard error of the mean (n=3). *P<0.05, **P<0.01 and ***P<0.001 vs. Cont group; #P<0.05 and ##P<0.01 vs. LPS group. TA, tannic acid; LPS, lipopolysaccharide; Cont, control; FITC, fluorescein isothiocyanate; PI, propidium iodide.
9). Tannic acid alleviates lipopolysaccharide‑induced H9C2 cell apoptosis by suppressing reactive oxygen species‑mediated endoplasmic reticulum stress. Molecular Medicine Reports, 2021 (PubMed: 34080663) [IF=3.4]

Application: WB    Species: Rat    Sample: H9C2 cells

Figure 2. TA alleviates the LPS-induced apoptosis of H9C2 cells. (A) Flow cytometry was used to determine cell apoptotic rate using Annexin V-FITC/PI staining. Apoptotic cells included Annexin V+/PI- and Annexin V+/PI+ cells. (B) Quantitative analysis of the apoptotic rate of cells treated with LPS, TA and TA + LPS. (C) Protein expression levels of Bax, Bcl-2, cleaved caspase-9, cleaved caspase-3 and β-actin in H9C2 cells treated with LPS, TA or TA + LPS were determined by western blot analysis. (D) Semi-quantification of the protein expression levels of Bax/Bcl-2, cleaved caspase-9 and cleaved caspase-3 in the LPS, TA and TA + LPS groups. Data are presented as the mean ± standard error of the mean (n=3). *P
10). Cleavage of semaphorin 4 C interferes with the neuroprotective effect of the semaphorin 4 C/Plexin B2 pathway on experimental intracerebral hemorrhage in rats. Journal of chemical neuroanatomy, 2023 (PubMed: 37482144) [IF=2.8]
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