Recombinant human active Caspase-3 protein (ab52314)

製品の概要

製品の詳細

  • 由来Recombinant
  • 由来Escherichia coli
  • アミノ酸配列
    • アクセッション番号P42574
    • 生物種Human

特性

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.

  • 生理活性

    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.

  • アプリケーション

    SDS-PAGE

  • 精製度> 90 % SDS-PAGE.

  • 製品の状態Lyophilised
  • 備考After 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...

前処理および保存

  • 保存方法および安定性

    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.

  • 再構成Reconstitute to 0.1 µg/µl in PBS containing 15% glycerol.

関連情報

  • 別名
    • 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
  • 機能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.
  • 組織特異性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.
  • 配列類似性Belongs to the peptidase C14A family.
  • 翻訳後修飾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.
  • 細胞内局在Cytoplasm.
  • Information by UniProt

Recombinant human active Caspase-3 protein 画像

  • 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.

Recombinant human active Caspase-3 protein (ab52314) 使用論文

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"