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Human/Mouse Caspase-3 Antibody

Catalog #: AF-605-NA
22 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF-605-NA
AF-605-SP
Detection of Human and Mouse Caspase‑3 by Western Blot.
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Product Details
Citations (22)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse Caspase-3 Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human Caspase-3.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Caspase-3
Met1-His277
Accession # AAA65015
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
Simple Western
5 µg/mL
See below
Immunoprecipitation
1 µg/106 cells
See below
Knockout Validated
Caspase‑3 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Caspase‑3 knockout HeLa cell line.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human and Mouse Caspase-3 antibody by Western Blot. View Larger

Detection of Human and Mouse Caspase‑3 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line and DA3 mouse myeloma cell line untreated (-) or treated (+) with 1 µg/mL Staurosporine (STS) for 12 hours. PVDF Membrane was probed with 0.5 µg/mL of Goat Anti-Human Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-605-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Caspase-3 precursor at approximately 34 kDa (as indicated) and Caspase-3 (cleaved) at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Immunoprecipitation Immunoprecipitation of Human Caspase‑3. View Larger

Immunoprecipitation of Human Caspase‑3. Jurkat human acute T cell leukemia cell line was treated with indicated concentrations of Staurosporine for 0 or 4 hours. Caspase‑3 was immunoprecipitated from lysates of 1‑2 x 106cells following incubation with 1 or 4 µg Goat Anti-Human Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-605-NA) overnight at 4 ºC. Caspase‑3-antibody complexes were absorbed using Protein G expressing Staph cells (Sigma). Immunoprecipitated Caspase‑3 was detected by Western blot using 0.5 µg/mL Goat Anti-Human Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑605‑NA). View our recommended buffer recipes for immunoprecipitation.

Simple Western Detection of Human Caspase-3 antibody by Simple Western<SUP>TM</SUP>. View Larger

Detection of Human Caspase‑3 by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, and Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Caspase-3 at approximately 40 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-605-NA) 1:50 dilution followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Knockout Validated Western Blot Shows Human Caspase-3 Antibody Specificity by Using Knockout Cell Line. View Larger

Western Blot Shows Human Caspase‑3 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Caspase-3 knockout HeLa cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human/Mouse Caspase-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-605-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Caspase-3 at approximately 32 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Caspase-3

Caspase-3 (Cysteine-aspartic acid protease 3/Casp3; also Yama, apopain and CPP32) is a 29 kDa member of the peptidase C14A family of enzymes (1, 2, 3). It is widely expressed and is an integral component of the apoptotic cascade. Caspase-3 is considered to be the major executioner caspase; that is, the primary downstream mediator of apoptotic-associated proteolysis (2, 3, 4). Active Caspase-3 is known to utilize a Cys residue to cleave multiple substrates, including PARP, proIL-16, PKC-gamma & -δ, procaspases 6, 7 and 9, and beta -catenin (1). Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein (5, 6, 7). Normally, it is an inactive, cytosolic homodimer, but following an upstream signal that activates processing proteases, procaspase-3 undergoes proteolytic cleavage (1, 2, 8, 9). This generates an N-terminal 175 aa p20/20 kDa subunit plus a 102 aa C-terminal p12/12 kDa subunit, followed by further processing of the p20 subunit at Asp28 to generate a final p17 subunit (aa 29-175) (9). The p17 and p12 subunits noncovalently heterodimerize, and subsequently associate with another p17/p12 heterodimer to form an active antiparallel homodimer. The p17 subunit contains the enzyme active site (aa 161-165), with an embedded catalytic Cys which is normally nitrosylated and inactive. Full activation requires both proteolytic processing and Cys163 denitrosylation (10). Multiple proteases can use Caspase-3 as a substrate including Caspase-6, -8, and -10, granzyme B, and Caspase-3 itself (9, 11, 12, 13). 

References
  1. Chowdhury, I. et al. (2008) Comp. Biochem. Physiol. B 151:10.
  2. Boatright, K.M. & G.S. Salvesen (2003) Curr. Opin. Cell Biol. 15:725.
  3. Launay, S. et al. (2005) Oncogene 24:5137.
  4. Walsh, J.G. et al. (2008) Proc. Natl. Scad. Sci. USA 105:12815.
  5. Nicholson, D.W. et al. (1995) Nature 376:37.
  6. Tewari, M. et al. (1995) Cell 81:801.
  7. Fernandes-Alnemri, T. et al. (1994) J. Biol. Chem. 269:30761.
  8. Milisav, I. et al. (2009) Apoptosis 14:1070.
  9. Han, Z. et al. (1997) J. Biol. Chem. 272:13432.
  10. Rossig, L. et al. (1999) J. Biol. Chem. 274:6823.
  11. Rank, K.B. et al. (2001) Protein Expr. Purif. 22:258.
  12. Atkinson, E.A. et al. (1998) J. Biol. Chem. 273:21261.
  13. Cohen, G.M. (1997) Biochem. J. 326:1.
Entrez Gene IDs
836 (Human); 12367 (Mouse)
Alternate Names
Apopain; apoptosis-related cysteine protease; CASP3; CASP-3; caspase 3, apoptosis-related cysteine peptidase; Caspase3; Caspase-3; CPP32; CPP-32; CPP32B; CPP32SREBP cleavage activity 1; Cysteine protease CPP32; EC 3.4.22; EC 3.4.22.56; LICE-1; PARP cleavage protease; procaspase3; Protein Yama; SCA-1; YAMA

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Citations for Human/Mouse Caspase-3 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

22 Citations: Showing 1 - 10
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  1. Hypoxia-Ischemia and Hypothermia Independently and Interactively Affect Neuronal Pathology in Neonatal Piglets with Short-Term Recovery.
    Authors: O'Brien, C E, Santos, P T Et al.
    Dev Neurosci
  2. Preclinical studies identify non-apoptotic low-level caspase-3 as therapeutic target in pemphigus vulgaris.
    Authors: Luyet C, Schulze K, Sayar B, Howald D, Muller E, Galichet A
    PLoS ONE, 2015-03-06;10(3):e0119809.
  3. Discovery of Small-Molecule Inhibitors of the HSP90-Calcineurin-NFAT Pathway against Glioblastoma
    Authors: Zhenzhen Liu, Hongli Li, Lian He, Yu Xiang, Chengsen Tian, Can Li et al.
    Cell Chemical Biology
  4. Enhanced Apoptosis and Loss of Cell Viability in Melanoma Cells by Combined Inhibition of ERK and Mcl-1 Is Related to Loss of Mitochondrial Membrane Potential, Caspase Activation and Upregulation of Proapoptotic Bcl-2 Proteins
    Authors: Z Peng, B Gillissen, A Richter, T Sinnberg, MS Schlaak, J Eberle
    International Journal of Molecular Sciences, 2023-03-04;24(5):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Toxicity Assessment of Curculigo orchioides Leaf Extract Using Drosophila melanogaster: A Preliminary Study
    Authors: S Kushalan, LC D'Souza, K Aloysius, A Sharma, S Hegde
    International Journal of Environmental Research and Public Health, 2022-11-18;19(22):.
    Species: Drosophila melanogaster
    Sample Types: Tissue Homogenates
    Applications: IHC
  6. MitoTEMPOL Inhibits ROS-Induced Retinal Vascularization Pattern by Modulating Autophagy and Apoptosis in Rat-Injected Streptozotocin Model
    Authors: Rova Virgana, Nur Atik, Julia Windi Gunadi, Evelyn Jonathan, Dona Erisa Ramadhani, Ray Sebastian Soetadji et al.
    Life (Basel)
  7. Potent pro-apoptotic combination therapy is highly effective in a broad range of cancers
    Authors: Antonella Montinaro, Itziar Areso Zubiaur, Julia Saggau, Anna-Laura Kretz, Rute M. M. Ferreira, Omar Hassan et al.
    Cell Death & Differentiation
  8. Activation of the executioner caspases-3 and -7 promotes microglial pyroptosis in models of multiple sclerosis
    Authors: BA McKenzie, JP Fernandes, MAL Doan, LM Schmitt, WG Branton, C Power
    J Neuroinflammation, 2020-08-29;17(1):253.
    Species: Human
    Sample Types: Protein, Tissue Homogenates, Whole Cells, Whole Tissue
    Applications: ICC, IHC, Western Blot
  9. Hypoxia-induced complement dysregulation is associated with microvascular impairments in mouse tracheal transplants
    Authors: MA Khan, T Shamma, S Kazmi, A Altuhami, HA Ahmed, AM Assiri, DC Broering
    J Transl Med, 2020-03-31;18(1):147.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  10. Apoptosis-mediated antiproliferation of A549 lung cancer cells mediated by Eugenia aquea leaf compound 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone and its molecular interaction with caspase receptor in molecular docking simulation
    Authors: YE Hadisaputr, N Cahyana, M Muchtaridi, R Lesmana, T Rusdiana, AY Chaerunisa, I Sufiawati, T Rostinawat, A Subarnas
    Oncol Lett, 2020-03-19;19(5):3551-3557.
    Species: Human
    Sample Types: Cell Lysate
    Applications: Western Blot
  11. Levels of caspase-3 and histidine-rich glycoprotein in the embryo secretome as biomarkers of good-quality day-2 embryos and high-quality blastocysts
    Authors: H Kaihola, FG Yaldir, T Bohlin, R Samir, J Hreinsson, H Åkerud
    PLoS ONE, 2019-12-19;14(12):e0226419.
    Species: Human
    Sample Types: Tissue Culture Supernates
    Applications: Proximity Extension Assay
  12. Lipid storage droplet protein 5 reduces sodium palmitate?induced lipotoxicity in human normal liver cells by regulating lipid metabolism?related factors
    Authors: X Ma, F Cheng, K Yuan, K Jiang, T Zhu
    Mol Med Rep, 2019-06-06;20(2):879-886.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  13. The Effect of Long-Term Administration of Fatty Acid Amide Hydrolase Inhibitor URB597 on Oxidative Metabolism in the Heart of Rats with Primary and Secondary Hypertension
    Authors: M Biernacki, W ?uczaj, I Jarocka-Ka, E Ambro?ewic, M Toczek, E Skrzydlews
    Molecules, 2018-09-14;23(9):.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  14. Overlooked benefits of using polyclonal antibodies
    Authors: Carl A Ascoli, Birte Aggeler
    BioTechniques
  15. TGF‑ beta 1 mediates the effects of aspirin on colonic tumor cell proliferation and apoptosis
    Authors: Yuyi Wang, Chi Du, Nan Zhang, Mei Li, Yanyang Liu, Maoyuan Zhao et al.
    Oncology Letters
  16. Bok is a genuine multi-BH-domain protein that triggers apoptosis in the absence of Bax and Bak and augments drug response
    Authors: S Einsele-Sc, S Malmsheime, K Bertram, D Stehle, J Johänning, M Manz, PT Daniel, BF Gillissen, K Schulze-Os, F Essmann
    J Cell Sci, 2016-04-13;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  17. Multiple Mechanisms Mediate Resistance to Sorafenib in Urothelial Cancer
    Authors: Judith Knievel, Wolfgang A. Schulz, Annemarie Greife, Christiane Hader, Tobias Lübke, Ingo Schmitz et al.
    International Journal of Molecular Sciences
  18. Overexpressing cellular repressor of E1A-stimulated genes protects mesenchymal stem cells against hypoxia- and serum deprivation-induced apoptosis by activation of PI3K/Akt.
    Authors: Deng J, Han Y, Yan C, Tian X, Tao J, Kang J, Li S
    Apoptosis, 2010-04-01;15(4):463-73.
    Species: Rat
    Sample Types: Cell Lysates
    Applications: Western Blot
  19. Ellipticine derivative NSC 338258 represents a potential new antineoplastic agent for the treatment of multiple myeloma.
    Authors: Tian E, Landowski TH, Stephens OW, Yaccoby S, Barlogie B, Shaughnessy JD
    Mol. Cancer Ther., 2008-03-04;7(3):500-9.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  20. Photochemically mediated delivery of AdhCMV-TRAIL augments the TRAIL-induced apoptosis in colorectal cancer cell lines.
    Authors: Engesaeter BØ, Bonsted A, Lillehammer T, Engebraaten O, Berg K, Maelandsmo GM
    Cancer Biol. Ther., 2006-11-19;5(11):1511-20.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  21. The B30.2 domain of pyrin, the familial Mediterranean fever protein, interacts directly with caspase-1 to modulate IL-1beta production.
    Authors: Chae JJ, Wood G, Masters SL, Richard K, Park G, Smith BJ, Kastner DL
    Proc. Natl. Acad. Sci. U.S.A., 2006-06-19;103(26):9982-7.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  22. Stenotrophomonas maltophilia Serine Protease StmPr1 Induces Matrilysis, Anoikis, and Protease-Activated Receptor-2 Activation in Human Lung Epithelial Cells
    Authors: AL DuMont, NP Cianciotto
    Infect. Immun., 2017-11-17;0(0):.

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Human/Mouse Caspase-3 Antibody
By Anonymous on 04/08/2022
Application: WB Sample Tested: A549 human lung carcinoma cell line Species: Human