Recombinant human active Caspase-3 protein (ab52314)

Overview

Description

  • NatureRecombinant
  • SourceEscherichia coli
  • Amino Acid Sequence
    • AccessionP42574
    • SpeciesHuman

Specifications

Our Abpromise guarantee covers the use of ab52314 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Biological activity

    Specific Activity:300,000 - 400,000 units/mg.

    Enzyme Assay: Use at 1 unit/assay. One unit of the recombinant caspase-3 is the enzyme activity that cleaves 1 nmol of the caspase substrate DEVD-pNA (pNA: pnitroanaline) per hour at 37degC in a reaction solution containing 50 mM Hepes, pH 7.2, 50 mM NaCl, 0.1% Chaps, 10 mM EDTA, 5% Glycerol, and 10 mM DTT.

  • Applications

    SDS-PAGE

  • Purity> 90 % SDS-PAGE.

  • FormLyophilised
  • Additional notesAfter initial reconstitution, aliquot product into individual tubes and immediately refreeze at –70degC.


    This recombinant active human caspase 3 expressed in E. coli spontaneously undergoes autoprocessing to yield subunits characteristic of the native enzyme. It can be useful in studying enzyme regulation, determining target substrates, screening caspase inhibitors, or as a positive control in caspase activity assays.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.

    Preservative: None
    Constituents: 15% Glycerol, PBS

    This product is an active protein and may elicit a biological response in vivo, handle with caution.

  • ReconstitutionReconstitute to 0.1 µg/µl in PBS containing 15% glycerol.

General Info

  • Alternative names
    • Apopain
    • CASP 3
    • CASP-3
    • CASP3
    • CASP3_HUMAN
    • Caspase 3
    • Caspase-3 subunit p12
    • CPP 32
    • CPP-32
    • CPP32B
    • Cysteine protease CPP32
    • PARP cleavage protease
    • Protein Yama
    • SCA-1
    • SCA1
    • SREBP cleavage activity 1
    • Yama
    see all
  • FunctionInvolved 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.
  • Tissue specificityHighly 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.
  • Sequence similaritiesBelongs to the peptidase C14A family.
  • 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.
  • Cellular localizationCytoplasm.
  • Information by UniProt

Recombinant human active Caspase-3 protein images

  • Lane 1 : Recombinant caspase 3 protein (Active) ab52314 at 2 ng. Lane 2 : Hela-Staurosporine ab52203 at 40 ug. Lane 3 : Jurkat-Staurosporine at 20 ug. Lane 4 : HeLa-Vehicle ab52203 at 40 ug. Lane 5 : Jurkat-Vehicle at 20 ug. Primary : Anti Caspase 3 (ab32351) 1:2000. Secondary : Goat polyclonal to Rabbit IgG ? H&L at 1:10000. Development: ECL with exposure 10 min. Predicted band size : 17 kDa. Migration of observed specific band is indicated with arrow
  • Lane 1 = Caspase 3 protein (Active) (ab52314) ? 20 ng. Lane 2 = Caspase 9 protein (Active) (ab52203) ? 20 ng. Lane 3 = Extract of HeLa cells treated with vehicle (ab136806) ? 20 ug. Lane 4 = Extract of HeLa cells treated with staurosporine (ab136806) ? 20 ug. SDS PAGE performed under reducing conditions (100mM DTT ? Sample heated at 50?C). Primary : Lanes 1-4: Anti active+pro Caspase 3 antibody (ab13585) at 1 ug/mL. Secondary : Lanes 1-4: Goat anti mouse IgG(H&L)-HRP at 1:10000. Development: ECL with 1 min exposure. Blocking: in 5% Milk + PBS overnight at 4 C. Primary antibody: in 5% Milk + PBS for 2 hours at RT. Secondary antibody: in 5% Milk + PBS for 2 hours at RT. Predicted band size : 17 kDa. Observed band size : 32 kDa and 17 kDa.

References for Recombinant human active Caspase-3 protein (ab52314)

ab52314 has not yet been referenced specifically in any publications.

Product Wall

Thank you for your enquiry.

Here is the buffer recipe:

Bicarbonate/carbonate coating buffer (100 mM)
Antigen or antibody should be diluted in coating buffer to immobilize them to the wells:
3.03 g Na2CO3
6.0 g NaHCO3 ...

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Thank you for your enquiry.

I have found that this active Caspase protein, ab52314, does contain both 17kDa and 12kDa subunits.

Here is a link to our sandwich ELISA protocol:
http://www.abcam.com/index.html?pageconfig=resource&...

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I am sorry but that information is propietary. I apologize for the inconvenience.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"