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  • Product Name
    Cleaved Caspase 3 Antibody
  • Catalog No.
    AF7022
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Hm,Ms,Rt
  • UniProt
  • Mol.Wt.
    17kDa
  • Concentration
    1mg/ml
  • Browse similar products>>

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

Reactivity:

Human,Mouse,Rat

Source:

Rabbit

Clonality:

Polyclonal

Purification:

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

Specificity:

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

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

Immunogen:

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

Uniprot:



>>Visit The Human Protein Atlas

Gene id:

Molecular Weight:

Observed Mol.Wt.: 17kDa.
Predicted Mol.Wt.: 32kDa.

Subcellular Location:

Cytoplasm.

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.

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:
        10         20         30         40         50
MENTENSVDS KSIKNLEPKI IHGSESMDSG ISLDNSYKMD YPEMGLCIII
60 70 80 90 100
NNKNFHKSTG MTSRSGTDVD AANLRETFRN LKYEVRNKND LTREEIVELM
110 120 130 140 150
RDVSKEDHSK RSSFVCVLLS HGEEGIIFGT NGPVDLKKIT NFFRGDRCRS
160 170 180 190 200
LTGKPKLFII QACRGTELDC GIETDSGVDD DMACHKIPVE ADFLYAYSTA
210 220 230 240 250
PGYYSWRNSK DGSWFIQSLC AMLKQYADKL EFMHILTRVN RKVATEFESF
260 270
SFDATFHAKK QIPCIVSMLT KELYFYH

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.

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 > TNF signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > MAPK signaling pathway.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Colorectal cancer.(View pathway)
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Cancers: Overview > MicroRNAs in cancer.
· Human Diseases > Neurodegenerative diseases > Parkinson's disease.
· Human Diseases > Cardiovascular diseases > Viral myocarditis.
· Human Diseases > Infectious diseases: Bacterial > Legionellosis.
· Human Diseases > Cancers: Specific types > Small cell lung cancer.(View pathway)
· Human Diseases > Infectious diseases: Bacterial > Pertussis.
· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).
· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.
· Human Diseases > Infectious diseases: Parasitic > Amoebiasis.
· Human Diseases > Cancers: Overview > Viral carcinogenesis.
· Human Diseases > Infectious diseases: Viral > Hepatitis B.
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Infectious diseases: Bacterial > Epithelial cell signaling in Helicobacter pylori infection.
· Human Diseases > Infectious diseases: Bacterial > Tuberculosis.
· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.
· Human Diseases > Neurodegenerative diseases > Huntington's disease.
· Human Diseases > Endocrine and metabolic diseases > Non-alcoholic fatty liver disease (NAFLD).
· Human Diseases > Neurodegenerative diseases > Alzheimer's disease.
· Human Diseases > Infectious diseases: Parasitic > Toxoplasmosis.
· Organismal Systems > Immune system > IL-17 signaling pathway.(View pathway)
· Organismal Systems > Immune system > Natural killer cell mediated cytotoxicity.(View pathway)
· Organismal Systems > Nervous system > Serotonergic synapse.

Western blot analysis of extracts from various samples, using Cleaved Caspase 3 Antibody. Lane 1: Hela treated with blocking peptide; Lane 2: Hela (etoposide treated, 25uM 5h); Lane 3: MCF7 (etoposide treated, 25uM 5h); Lane 4: Mouse heart; Lane 5: Mouse spleen.
Purkinje Cell Degeneration and Motor Coordination Deficits in a New Mouse Model of Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay Man Ding, Chao Weng, Shanghua Fan, Qian Cao, and Zuneng Lu*
BCL2L10 inhibits growth and metastasis of hepatocellular carcinoma both in vitro and in vivo Y Bai, J Wang, J Han, XL Xie, CG Ji… Molecular …, 2016 Wiley Online Library
This image is a courtesy of anonymous review.
Western blot analysis of extracts from COLO(staurosporine treated) cells, using Caspase 3 (p17,Cleaved-Asp175) Antibody. The lane on the left is treated with the synthesized peptide.
AF7022 at 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
Immunohistochemical staining of formalin fixed paraffin-embedded ehrlich carcimoma tissue using c-cadpase3 antibody at 1:100 diluting
AF7022 staining Hela by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25°C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37°C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.

Reference Citations:

1). Li R et al. Transcription factor 3 controls cell proliferation and migration in glioblastoma multiforme cell lines. Biochem Cell Biol 2016 Jun;94(3):247-55 (PubMed: 27105323)

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


2). Chen F et al. A ROS-mediated lysosomal-mitochondrial pathway is induced by ginsenoside Rh2 in hepatoma HepG2 cells. Food Funct 2015 Dec;6(12):3828-37 (PubMed: 26449932)

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.


3). Chen W et al. RhoB Acts as a Tumor Suppressor That Inhibits Malignancy of Clear Cell Renal Cell Carcinoma. PLoS One 2016 Jul 6;11(7):e0157599 (PubMed: 27384222)

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


4). Ding M et al. Purkinje Cell Degeneration and Motor Coordination Deficits in a New Mouse Model of Autosomal Recessive Spastic Ataxia of Charlevoix-Saguenay. Front Mol Neurosci 2017 May 1;10:121 (PubMed: 28588446)

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

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


5). Mengyang Zhao et al. Packaging cordycepin phycocyanin micelles for the inhibition of brain cancer. J. Mater. Chem. B 2017;5:6016-6026

6). Song Y et al. AMPK activation-dependent autophagy compromises oleanolic acid-induced cytotoxicity in human bladder cancer cells. Oncotarget 2017 Jul 4;8(40):67942-67954 (PubMed: 28978086)

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


7). Zhang X et al. Neuroprotective Effect of Modified Xijiao Dihuang Decoction against Oxygen-Glucose Deprivation and Reoxygenation-Induced Injury in PC12 Cells: Involvement of TLR4-MyD88/NF-κB Signaling Pathway. Evid Based Complement Alternat Med 2017;2017:3848595 (PubMed: 29234386)

8). Wu D et al. Olaquindox disrupts tight junction integrity and cytoskeleton architecture in mouse Sertoli cells. Oncotarget 2017 Aug 16;8(51):88630-88644 (PubMed: 29179463)

9). Yin L et al. Bacillus spore-based oral carriers loading curcumin for the therapy of colon cancer. J Control Release 2018 Feb 10;271:31-44 (PubMed: 29274436)

10). 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)

11). Zheng Z et al. The co-treatment of metformin with flavone synergistically induces apoptosis through inhibition of PI3K/AKT pathway in breast cancer cells. Oncol Lett 2018 Apr;15(4):5952-5958 (PubMed: 29552226)

12). Liu Y et al. Increased autophagy in EOC re-ascites cells can inhibit cell death and promote drug resistance. Cell Death Dis 2018 Apr 1;9(4):419 (PubMed: 29549251)

13). Liang Q et al. The role of IP3R-SOCCs in Cr(vi)-induced cytosolic Ca2+ overload and apoptosis in L-02 hepatocytes. Toxicol Res (Camb) 2018 Apr 26;7(3):521-528 (PubMed: 30090602)

14). 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)

15). 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)

16). 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)

17). 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)

18). 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)

19). Sun C et al. Hsa-miR-326 targets CCND1 and inhibits non-small cell lung cancer development. Oncotarget 2016 Feb 16;7(7):8341-59 (PubMed: 26840018)

Application: WB 1/1000    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.


20). Feng X et al. Graphene oxide induces p62/SQSTM-dependent apoptosis through the impairment of autophagic flux and lysosomal dysfunction in PC12 cells. Acta Biomater 2018 Sep 28 (PubMed: 30273743)

21). Xu J et al. Inhibiting succinate dehydrogenase by dimethyl malonate alleviates brain damage in a rat model of cardiac arrest. Neuroscience 2018 Oct 6 (PubMed: 30300703)

22). Feng Z et al. FTY720 attenuates intestinal injury and suppresses inflammation in experimental necrotizing enterocolitis via modulating CXCL5/CXCR2 axis. Biochem Biophys Res Commun 2018 Nov 10;505(4):1032-1037 (PubMed: 30314693)

23). Yu G et al. The protective effect of low-energy shock wave on testicular ischemia-reperfusion injury is mediated by the PI3K/AKT/NRF2 pathway. Life Sci 2018 Oct 12 (PubMed: 30321543)

24). Shang J et al. CircPAN3 mediates drug resistance in acute myeloid leukemia through the miR-153-5p/miR-183-5p-XIAP axis. Exp Hematol 2018 Nov 2 (PubMed: 30395908)

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Catalog Number :

AF7022-BP

Price/Size :

$200/1mg.
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 1 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.

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.