Overview

  • Product nameCalpain Activity Assay试剂盒
  • Tests
    100 x 1 assay
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeQuantitative
  • Assay time
    2h 00m
  • Product overview

    Abcam's Calpain Activity Assay试剂盒provides optimized缓冲器and试剂for a convenient measurement of calpain activity. The Extraction缓冲器provided with the试剂盒specifically extracts cytosolic proteins without contaminations of cell膜and lysosome proteases. The Extraction缓冲器also prevents auto-activation of calpain during the extraction procedure. Thus, the试剂盒detects only activated calpain in cytosol upon treatment of cells with inducers (e.g., chemicals or drugs). The fluorometric assay is based on the detection of cleavage of calpain substrate Ac-LLY-AFC. Ac-LLY-AFC emits blue light (λmax = 400 nm); upon cleavage of the substrate by calpain, free AFC emits a yellow-green fluorescence (λmax = 505 nm), which can be quantified using a fluorometer or a fluorecence plate reader. Comparison of the fluorescence intensity from a treated sample with a normal control allows determination of the changes in calpain activity.


    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Activation of calpain is involved in many forms of physiological and pathological processes (e.g., apoptosis). Calpain activation requires cell membrane and Ca²+, and activated calpain is released into cytosol.

     

    If additional Ac-LLY-AFC substrate is needed, it can be purchased separately as ab171379.

  • Tested applicationsFunctional Studiesmore details

Properties

  • Storage instructionsPlease refer to protocols.
  • Components Identifier 100 tests
    10X Reaction Buffer Clear 1 x 1.5ml
    Active Calpain I (Positive Control) Green 1 x 10µl
    Calpain Inhibitor Z-LLY-FMK Orange 1 x 10µl
    Calpain Substrate Ac-LLY-AFC Amber 1 x 500µl
    Extraction Buffer WM 1 x 25ml
  • Research Areas
  • FunctionCalcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
  • Tissue specificityUbiquitous.
  • Sequence similaritiesBelongs to the peptidase C2 family.
    Contains 1 calpain catalytic domain.
    Contains 4 EF-hand domains.
  • Cellular localizationCytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding.
  • Information by UniProt
  • Alternative names
    • Calcium activated neutral proteinase 1
    • Calcium-activated neutral proteinase 1
    • Calpain 1 (mu/I) large subunit
    • Calpain 1 catalytic subunit
    • Calpain 1 large subunit
    • Calpain large polypeptide L1
    • Calpain mu type
    • Calpain mu-type
    • Calpain-1 catalytic subunit
    • Calpain-1 large subunit
    • CAN1_HUMAN
    • CANP
    • CANP 1
    • CANP1
    • CANPL1
    • CAPN 1
    • CAPN1
    • Cell proliferation inducing gene 30 protein
    • Cell proliferation inducing protein 30
    • Cell proliferation-inducing gene 30 protein
    • Micromolar calpain
    • Micromolar-calpain
    • muCANP
    • muCL
    see all

Applications

Our Abpromise guarantee covers the use of ab65308 in the following tested applications.

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

Application Abreviews Notes
Functional Studies Use at an assay dependent concentration.

Calpain Activity Assay Kit images

  • Typical Data for ab65308: Active Calpain (1 µg) was incubated at 37 °C for one hour using the Calpain Substrate with or without 20 µM Calpain Inhibitor.

Protocols

References for Calpain Activity Assay Kit (ab65308)

This product has been referenced in:

See all 4 Publications for this product

Product Wall


For the Calpain assay we highly recommend using fresh tissues or tissue samples snap-frozen after collection. The frozen tissue can be ground and then homogenized in the extraction buffer.

One point to note is : Inactive calpain remains a...

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The success of this kit with frozen samples depends on how well the samples were frozen, for how long they had been frozen, if they underwent repeated freeze thaw and so on.

Immediate freezing of the sample in aliquots and storing ...

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This kit has the substrate bound to the AFC fluorophore.

The success of this kit with frozen samples depends on how well the samples were frozen, for how long they had been frozen, if they underwent repeated freeze thaw and so on.
Immediate freezing of the sample in aliquots and storing for up to 2 mont...

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I can confirm that for the sample preparation the cells will not need to be trypsinized. They can be scraped from the flask after after adding the extraction buffer and incubated, but keep in mind that this would dilute the sample for the activity assa...

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This Calpain Activity Assay Kit includes Active Calpain I as positive control.

The Calpain protein provided in ab65308 is a native protein. We pack 14.5 ug of Calpain/tube in this kit.

Thank you once again for your enquiries.

I have obtained some further information for you.

Unfortunately, I cannot disclose the contents of the extraction buffer due to proprietary restrictions, but I can assure you that it is str...

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Hello. No, this is not exactly the same, the protein ab91019 comes from another source. They are very similar though. Please let me know if you have any further questions.

Thank you for your email.

The kit is specific to all forms of Calpain enzymes not only the calpain 1.

Biochemically this kit is based on the fact that the Calpain enzymes present in cell lysates cleave the calpain substrate Ac-LLY-...

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