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  • Product Name
    Cleaved Caspase 3 Antibody
  • Catalog No.
    AF7022
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity:
    Human, Mouse, Rat
  • Prediction:
    Pig(100%), Bovine(100%), Horse(86%), Sheep(100%), Rabbit(86%), Dog(100%), Xenopus(86%)
  • 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

Predicted Reactivity:

Pig(100%), Bovine(100%), Horse(86%), Sheep(100%), Rabbit(86%), Dog(100%), Xenopus(86%)

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 293t, using Cleaved Caspase 3 Antibody. The lane on the left was treated with blocking peptide.
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.
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 dilution of 1:100.
AF7022 staining Hela cells by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25°C. Samples were then incubated with primary Ab(AF7022 1:200) and mouse anti-beta tubulin Ab(T0023 1:200) for 1 hour at 37°C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.

Reference Citations:

1). 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) [IF=7.877]

2). 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) [IF=6.383]

3). 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) [IF=5.638]

4). Ding Q et al. The role of the apoptosis-related protein BCL-B in the regulation of mitophagy in hepatic stellate cells during the regression of liver fibrosis. Exp Mol Med 2019 Jan 11;51(1):6 (PubMed: 30635551) [IF=5.584]

5). 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)

6). Shen YH et al. Ethyl Rosmarinate Protects High Glucose-Induced Injury in Human Endothelial Cells. Molecules 2018 Dec 19;23(12) (PubMed: 30572638)

7). Chen WS et al. Investigation of Dacomitinib on Reducing Cell Necrosis and Enhancing Cell Apoptosis in C6 Glioma Rat Model by MRI. Biosci Rep 2019 Feb 19 (PubMed: 30782784)

8). Tu X et al. Pretreatment of Grape Seed Proanthocyanidin Extract Exerts Neuroprotective Effect in Murine Model of Neonatal Hypoxic-ischemic Brain Injury by Its Antiapoptotic Property. Cell Mol Neurobiol 2019 May 30 (PubMed: 31147852)

9). 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.


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). 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    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.


12). Zhang W et al. MicroRNA-124 expression in the brains of rats during early cerebral ischemia and reperfusion injury is associated with cell apoptosis involving STAT3. Exp Ther Med 2019 Apr;17(4):2870-2876 (PubMed: 30906474)

13). 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)

14). Xie J et al. 5-aminolevulinic acid photodynamic therapy reduces HPV viral load via autophagy and apoptosis by modulating Ras/Raf/MEK/ERK and PI3K/AKT pathways in HeLa cells. J Photochem Photobiol B 2019 Mar 21;194:46-55 (PubMed: 30925276)

15). 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.


16). 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)

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). Zhai Z et al. Andrographolide prevents human breast cancer-induced osteoclastic bone loss via attenuated RANKL signaling. Breast Cancer Res Treat 2014 Feb;144(1):33-45 (PubMed: 24481680)

19). 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)

20). Zhang W et al. MicroRNA-124 expression in the brains of rats during early cerebral ischemia and reperfusion injury is associated with cell apoptosis involving STAT3. Exp Ther Med 2019 Apr;17(4):2870-2876 (PubMed: 30906474)

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

22). 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)

23). 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)

24). Xiao Y et al. Clusterin increases mitochondrial respiratory chain complex I activity and protects against hexavalent chromium-induced cytotoxicity in L-02 hepatocytes. Toxicol Res (Camb) 2018 Nov 15;8(1):15-24 (PubMed: 30713657)

25). Chen D et al. Quxie Capsule Inhibits Colon Tumor Growth Partially Through Foxo1-Mediated Apoptosis and Immune Modulation. Integr Cancer Ther 2019 Jan-Dec;18:1534735419846377 (PubMed: 31030593)

26). 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


27). 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)

28). 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)

29). 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)

30). 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)

31). Wang L et al. Development of anisamide-targeted PEGylated gold nanorods to deliver epirubicin for chemo-photothermal therapy in tumor-bearing mice. Int J Nanomedicine 2019 Mar 8;14:1817-1833 (PubMed: 30880982)

32). 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


33). 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


34). 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)

35). 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)

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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 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%
Sheep
100%
Dog
100%
Bovine
100%
Rabbit
86%
Xenopus
86%
Horse
86%
Chicken
71%
Zebrafish
0%
High similarity Medium similarity Low similarity No similarity
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.