Recombinant human Cleaved Caspase-3 protein (Active) (ab52101)

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Key features and details

  • Expression system: Escherichia coli
  • Purity: > 95% SDS-PAGE
  • Active: Yes
  • Suitable for: Functional Studies

Description

  • Product name

    Recombinant human Cleaved Caspase-3 protein (Active)
    See all Cleaved Caspase-3 proteins and peptides

  • Biological activity

    SPECIFIC ACTIVITY: 300,000-400,000 units/mg

    UNIT DEFINITION: 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 37oC 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.

  • Purity

    > 95 % SDS-PAGE.

  • Expression system

    Escherichia coli

  • Accession

    P42574

  • Protein length

    Full length protein

  • Animal free

    No

  • Nature

    Recombinant

    • Species

      Human

    • Sequence

      MENTENSVDS KSIKNLEPKI IHGSESMDSG ISLDNSYKMD YPEMGLCIII NNKNFHKSTG MTSRSGTDVD AANLRETFRN LKYEVRNKND LTREEIVELM RDVSKEDHSK RSSFVCVLLS HGEEGIIFGT NGPVDLKKIT NFFRGDRCRS LTGKPKLFII QACRGTELDC GIETDSGVDD DMACHKIPVE ADFLYAYSTA PGYYSWRNSK DGSWFIQSLC AMLKQYADKL EFMHILTRVN RKVATEFESF SFDATFHAKK QIPCIVSMLT KELYFYH

    • Amino acids

      1 to 277

    • Additional sequence information

      (Gene ID 836)

Specifications

Our Abpromise guarantee covers the use of ab52101 in the following tested applications.

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

  • Applications

    Functional Studies

  • Form

    Lyophilized

  • Additional notes

    The recombinant active human Caspase 3 spontaneously undergoes autoprocessing to yield subunits characteristic of the native enzyme.

    Cleaved Caspase 3 is useful in studying enzyme regulation, determining target substrates, screening caspase inhibitors, or as a positive control in caspase activity assays. The active Caspase 3 preferentially cleaves Caspase 3 substrates e.g. DEVD-AFC or DEVD-pNA.

    MW: large (17 kD) and small (11 kD) subunits.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

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

    Constituents: PBS, 5% Glycerol (glycerin, glycerine), 1.71% HEPES, 1.71% Sodium chloride, 0.1% CHAPS, 0.16% EDTA, 0.12% DTT

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

  • Reconstitution
    Centrifuge the vial prior to opening. Reconstitute to 1 unit per µl in water.

General Info

  • Alternative names

    • active caspase 3
    • 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

  • 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.

  • 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.

  • Sequence similarities

    Belongs 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 localization

    Cytoplasm.
  • Information by UniProt

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (2)

Publishing research using ab52101? Please let us know so that we can cite the reference in this datasheet.

ab52101 has been referenced in 2 publications.

  • Yozwiak CE  et al. Toward a Microparticle-Based System for Pooled Assays of Small Molecules in Cellular Contexts. ACS Chem Biol 13:761-771 (2018). PubMed: 29365249
  • Cao L  et al. Pseudo-phosphorylation at AT8 epitopes regulates the tau truncation at aspartate 421. Exp Cell Res 370:103-115 (2018). PubMed: 29908160

Customer reviews and Q&As

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Question

How much protein is provided with the active Caspase 3 protein (Active) (ab52101)

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Answer

The active Caspase 3 protein (Active), ab52101, is provided at the equivalent of 100 units. There are between 300,000-400,000 units/mg. Therefore a total of between 0.25-0.33 ug of protein is provide per vial.

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Question

I ordered an active caspase3 protein ( Active) (ab52101). And I would like to know after I reconstruct the product with PBS solution, what components I have? Procaspase-3 ? Large unit of caspase-3 ( 17KDa) ? or small unit of caspase-3 ( 12KDa)?

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Answer

You have received the recombinant full length protein: processed form of Caspase 3 consists of large (17kDa) and small (12kDa) subunits which associate to form an active enzyme.

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Question

Purchased the protein and reconstituted as recommended, used the protein successfully during first experiment. Stored the protein as directed, but when went back to use it 2 weeks later, the protein was not active. Have tried a total 4 times, using varying amounts of protein, but still no signal. Protein may have degraded.

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Answer

Thank you for calling Abcam earlier today.

I am sorry about the issues you have been having with the Caspase 3 protein. As we discussed, I have requested that you be sent (via Cedarlane) a free of charge replacement for the protein.

If there is anything else I can help you with, please let me know.

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Question


Our lab want to purchase caspase 1 (ab39901) and caspase 3 (ab52101) for
my project. We have one question regarding these products. We want to know
if we are going to cut our protein (40ug) with caspase 1 or 3, how many ul of
caspase 1 or 3 we need to add into the buffer in order to have a useful reaction?
Another question is I checked your website but I only found 25 units in caspase 1
and 100 units in caspase 3. Can I have the information about how many ul or ml
you have in one tube of caspase 1 or 3?
Thank you for all the information.

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Answer

Thank you for contacting Abcam.

For ab39901, Caspase 1, we recommend using 1 unit/assay for analyzing caspase activity and this protein has a biological activity : >5000 units/mg and it is supplied lyophilized and it should be reconstituted to 1 unit per µl in PBS containing 15% glycerol (and as it supplied as 25 units you would end up with 25ul of solution).

For ab52101, Caspase 3, 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 37oC 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. You are supplied with 100 units and again it is lyophilized on arrival an you should reconstitute to 1 unit per µl in PBS containing 15% glycerol (therefore you would have 100ul of solution).

The above are recommended starting dilutions, however the optimal dilutions/concentrations should be determined by the end user.

Please let me know if there is anything else I can help you with.

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Question

1) can enzyme be active in other solutions, such as PBS?
2) How long is the enzyme active when put at 37 degrees in PBS, without substrate?
3) How long is the enzyme active when put at 37 degrees inthe buffer 50 mM Hepes, pH 7.2, 50 mM NaCl, 0.1% Chaps, 10 mM EDTA, 5% Glycerol, and 10 mM DTT, without substrate?

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Answer

Thank you for contacting us.

Here are the answers to your questions sent bythe lab:

1. We have not specifically tested the activity of this enzyme in PBS and hence we will not be able to comment on this. The enzyme is definitely stable in PBS, however as indicated in the data sheet it will be optimally active only 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. These components are required for the enzyme to be active on the substrate.

2. How long is the enzyme active when put at 37 degrees in PBS, without substrate? - We do not have this data.

3. Without the substrate when the enzyme is in buffer 50 mM Hepes, pH 7.2, 50 mM NaCl, 0.1% Chaps, 10 mM EDTA, 5% Glycerol, and 10 mM DTT, the protein should be active for 1-2 hrs.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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