Human Caspase-3 Antibody

  (6 citations)
(1 Review)
    
Datasheet / CoA / SDS
Product Details
Citations (6)
FAQs
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  • Species Reactivity
    Human
  • Specificity
    Detects human precursor Caspase-3 and the 18 kDa subunit generated during apoptosis.
  • Source
    Monoclonal Mouse IgG2B Clone # 84803
  • Purification
    Protein A or G purified from ascites
  • Immunogen
    E. coli-derived recombinant human Caspase-3 (18 kDa subunit)
    aa 29-175
    Accession # P42574
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    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.
Data Examples
Detection of Human Precursor Caspase‑3 and p18 Subunit by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 1 μM staurosporine (STS) for 3 hours. PVDF membrane was probed with 1 µg/mL of Human Caspase‑3 Monoclonal Antibody (Catalog # MAB707), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for  precursor Caspase‑3 and the p18 subunit at approximately 36 and 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Knockout Validated
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 2 µg/mL of Mouse Anti-Human Caspase‑3 Monoclonal Antibody (Catalog # MAB707) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
    Reconstitution Buffer Available
  • 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 heterodimer that belongs to the peptidase C14A family of enzymes. It is widely expressed, and considered to be the major executioner caspase in the apoptotic cascade. Human procaspase-3 is a 32 kDa, 277 amino acid (aa) protein and is normally an inactive homodimer. Following cell stress/activation, procaspase-3 undergoes proteolysis to generate an N-terminal 148 aa p17/17 kDa subunit (aa 29-175), plus a 102 aa C-terminal p12/12 kDa subunit. These subunits noncovalently heterodimerize, and associate with another p17/p12 heterodimer to form an active enzyme. There is one potential variant that shows an alternative start site nine aa upstream of the standard start site coupled with a 21 aa substitution for aa 162-277. Over aa 29-175, human and mouse caspase-3 share 87% aa identity.

 

  • 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
Related Research Areas
Citations:

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.

6 Citations: Showing 1 - 6
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Species
Applications
Sample Type
  1. Preeclampsia is associated with alterations in the p53-pathway in villous trophoblast.
    Authors: Sharp A, Heazell A, Baczyk D, Dunk C, Lacey H, Jones C, Perkins J, Kingdom J, Baker P, Crocker I
    PLoS ONE, 2014;9(1):e87621.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: WB
  2. PMA synergistically enhances apicularen A-induced cytotoxicity by disrupting microtubule networks in HeLa cells.
    Authors: Seo K, Kim J, Park J, Song K, Yun E, Park J, Kweon G, Yoon W, Lim K, Hwang B
    BMC Cancer, 2014;14(0):36.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Interactions between important regulatory proteins and human alphaB crystallin.
    Authors: Ghosh JG, Shenoy AK, Clark JI
    Biochemistry, 2007;46(21):6308-17.
    Species: Human
    Sample Type: Recombinant Protein
    Application: ELISA-based Protein Pin Array
  4. Developmental differences in the responses of IL-6 and IL-13 transgenic mice exposed to hyperoxia.
    Authors: Choo-Wing R, Nedrelow JH, Homer RJ, Elias JA, Bhandari V
    Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(1):L142-50.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  5. Proteolytically processed soluble tumor endothelial marker (TEM) 5 mediates endothelial cell survival during angiogenesis by linking integrin alpha(v)beta3 to glycosaminoglycans.
    Authors: Vallon M, Essler M
    J. Biol. Chem., 2006;281(45):34179-88.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Inhibition of lymphotoxin-beta receptor-mediated cell death by survivin-DeltaEx3.
    Authors: You RI, Chou YC
    Cancer Res., 2006;66(6):3051-61.
    Species: Human
    Sample Type: Cell Lysates
    Application: IP

FAQs

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Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG2B Isotype Control

Ctrl MAB004 34  
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Mouse IgG2B Isotype Control

Ctrl MAB0041 7  
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Mouse IgG2B Isotype Control

Ctrl MAB0042 6
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Reconstitution Buffers
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Reconstitution Buffer 1 (PBS)

RB01
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 32  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 12  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow , IHC , ICC NL007 10
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow , IHC , ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 4  
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG Antibody

WB AF007 4
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow , IHC , ICC NL009 5
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rat Anti-Mouse IgG2B APC-conjugated Antibody

Flow F0133 1  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Rat Anti-Mouse IgG2B PE-conjugated Antibody

Flow F0132  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Rat Anti-Mouse IgG2B Fluorescein-conjugated Antibody

Flow F0134 1
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Mouse IgG2B Antibody

Flow MAB0043  
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Average Rating: 5 (Based on 1 review)

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Read what people are saying who have used Human Caspase-3 Antibody.

We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Western Blot Human Caspase-3 Antibody MAB707 Array MAB707
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Excellent
 WB Human Anonymous 02/28/2017
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Western Blot Human Caspase-3 Antibody MAB707
Western Blot: Human Caspase-3 Antibody [MAB707].

Summary

Application Western Blot
Sample Tested Blood erythrocytes
Species Human

Other Experimental Details

Other Experimental Details The antibody works fine for what it is, but I think it will not work for our experiments. Compared to Jurkat or other cell types you only get very faint bands for caspase in RBCs unless you fix your membrane after the transfer, this gave a false positive comparing a negative and positive control for caspase activation. We had to switch to measuring samples only with flow. If your experiments are compatible it's a good antibody though. We tried this experiment with several antibodies of different companies and this one did give a nicer looking band.

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