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  • Product Name
    Cleaved-Caspase 3 (Asp175), p17 Antibody
  • Catalog No.
    AF7022
  • RRID
    AB_2835326
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA(peptide)
  • Reactivity
    Human, Mouse, Rat, Bovine
  • Prediction
    Pig, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus
  • UniProt
  • Mol.Wt
    17kD;
    32kDa(Calculated).
  • Concentration
    1mg/ml
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Product Information

Alternative Names:Expand▼

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;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.

Reactivity:

Human, Mouse, Rat, Bovine

Predicted Reactivity:

Pig, Bovine, Horse, Sheep, Rabbit, Dog, Xenopus

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).

Specificity:

Cleaved-Caspase 3 (Asp175,p17) Antibody detects endogenous levels of fragment of activated Caspase 3 resulting from cleavage adjacent to Asp175.

RRID:

AB_2835326
Please cite this product as: Affinity Biosciences Cat# AF7022, RRID:AB_2835326.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 13 months from date of receipt.

Immunogen Information in 3D

Immunogen:

The antiserum was produced against synthesized peptide derived from human Caspase 3.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

CASP3

Molecular Weight:

Observed Mol.Wt.: 17kD.
Predicted Mol.Wt.: 32kDa(Calculated)..

Subcellular Location:

Cytoplasm.

Tissue Specificity:

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.

Description:

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.

Sequence:
MENTENSVDSKSIKNLEPKIIHGSESMDSGISLDNSYKMDYPEMGLCIIINNKNFHKSTGMTSRSGTDVDAANLRETFRNLKYEVRNKNDLTREEIVELMRDVSKEDHSKRSSFVCVLLSHGEEGIIFGTNGPVDLKKITNFFRGDRCRSLTGKPKLFIIQACRGTELDCGIETDSGVDDDMACHKIPVEADFLYAYSTAPGYYSWRNSKDGSWFIQSLCAMLKQYADKLEFMHILTRVNRKVATEFESFSFDATFHAKKQIPCIVSMLTKELYFYH

Research Background

Function:

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.

Post-translational Modifications:

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.

Subcellular Location:

Cytoplasm.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Tissue Specificity:

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.

Subunit Structure:

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.

Similarity:

Belongs to the peptidase C14A family.

Research Fields

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 > 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.

Reference Citations:

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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). Li X et al. Cyanidin-3-O-glucoside restores spermatogenic dysfunction in cadmium-exposed pubertal mice via histone ubiquitination and mitigating oxidative damage. J Hazard Mater 2019 Nov 17:121706 (PubMed: 31796358) [IF=9.038]

3). Zhou K;Zheng Z;Li Y;Han W;Zhang J;Mao Y;Chen H;Zhang W;Liu M;Xie L;Zhang H;Xu H;Xiao J; et al. TFE3, a potential therapeutic target for Spinal Cord Injury via augmenting autophagy flux and alleviating ER stress. Theranostics 2020 Jul 23;10(20):9280-9302. (PubMed: 32802192) [IF=8.579]

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19). Li Y;Wu Y;Jiang K;Han W;Zhang J;Xie L;Liu Y;Xiao J;Wang X; et al. Mangiferin Prevents TBHP-Induced Apoptosis and ECM Degradation in Mouse Osteoarthritic Chondrocytes via Restoring Autophagy and Ameliorates Murine Osteoarthritis. Oxid Med Cell Longev 2019 Oct 15;2019:8783197. (PubMed: 31885823) [IF=5.076]

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Application: WB    Species:human;    Sample:HepG2

Fig. 5 Involvement of caspases in apoptosis induced by GRh2. (A) After incubating GRh2-loaded cells with or without 1 μM of Leu. Caspase activity was determined as indicated in section 2. Enzymatic activity is expressed as fold induction with respect to non-treated cells and represents the mean values ± SD. of three different experiments. *p < 0.001 significantly different from the control; #p < 0.001 significantly different from the cells treated with GRh2 in the absence of Leu. (B) Western blot assays of active fragments of caspase-3 and -9. The results from one representative experiment are shown. (C) The histogram represents quantification of caspase-3 and caspase-9 protein expression levels in GRh2 stimulated HepG2 cell samples using ImageJ64 software (levels of control cells/β-actin defined as 1). Results are presented as mean ± S.D. with triplicate measurement. *P < 0.01 vs. the control group.


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Application: WB    Species:human;    Sample:A172 cells

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


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77). Zhao T et al. Ligustrazine suppresses neuron apoptosis via the Bax/Bcl-2 and caspase-3 pathway in PC12 cells and in rats with vascular dementia. IUBMB Life 2018 Jan;70(1):60-70 (PubMed: 29247598) [IF=3.244]

78). Zhao T et al. Ligustrazine suppresses neuron apoptosis via the Bax/Bcl-2 and caspase-3 pathway in PC12 cells and in rats with vascular dementia. IUBMB Life 2018 Jan;70(1):60-70 (PubMed: 29247598) [IF=3.244]

79). Zhao T et al. Ligustrazine suppresses neuron apoptosis via the Bax/Bcl-2 and caspase-3 pathway in PC12 cells and in rats with vascular dementia. IUBMB Life 2018 Jan;70(1):60-70 (PubMed: 29247598) [IF=3.244]

80). Zhao T et al. Ligustrazine suppresses neuron apoptosis via the Bax/Bcl-2 and caspase-3 pathway in PC12 cells and in rats with vascular dementia. IUBMB Life 2018 Jan;70(1):60-70 (PubMed: 29247598) [IF=3.244]

81). Zhao T et al. Ligustrazine suppresses neuron apoptosis via the Bax/Bcl-2 and caspase-3 pathway in PC12 cells and in rats with vascular dementia. IUBMB Life 2018 Jan;70(1):60-70 (PubMed: 29247598) [IF=3.244]

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Application: WB    Species:human;    Sample:Not available


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Application: WB    Species:human;    Sample:U251 and A172 cells

Fig. 5. Expression levels of the caspase-3 and cleaved caspase-3 protein in U251 and A172 cells. (A, C) The results of western blots showing the levels of caspase-3 and cleaved caspase-3 protein increased after TCF3 shRNA treatments in both cells. (B, D) Histograms showing quantization of western blot and the data was normalized against β-actin. Results of at least three independent experiments. *p<0.05, **p


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110). Zhang W;Huang J;Tang Y;Yang Y;Hu H; et al. Inhibition of Fatty Acid Synthase (FASN) Affects the Proliferation and Apoptosis of HepG2 Hepatoma Carcinoma Cells via the β-catenin/C-myc Signaling Pathway. Ann Hepatol 2020 Mar 31;S1665-2681(20)30027-2. (PubMed: 32536483)

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Application: WB    Species:human;    Sample:Not available


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Application: WB    Species:Not available;    Sample:Not available

Figure 8: Ectopic expression of miR-326 promotes apoptosis in A549 and SPC-A-1 cells. (C) Western-blot of Bcl2 protein in A549 and SPC-A-1 cells after transfection.


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Catalog Number :

AF7022-BP
(Blocking peptide available as AF7022-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Pig
100%
Bovine
100%
Sheep
100%
Dog
100%
Horse
86%
Xenopus
86%
Rabbit
86%
Chicken
71%
Zebrafish
0%
High similarity Medium similarity Low similarity No similarity
P42574 as Substrate
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
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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