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MAK176

Sigma-Aldrich

Granzyme B Activity Assay Kit

Sufficient for 100 Fluorometric tests

Synonym(s):

CTLA1, GZMB Activity Assay Kit

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About This Item

EC Number:
200-664-3
UNSPSC Code:
12161503
NACRES:
NA.84

usage

sufficient for 100 fluorometric tests

detection method

fluorometric

relevant disease(s)

hematological disorder; immunological diseases; cancer

storage temp.

−20°C

Gene Information

human ... GZMB(3002)
mouse ... GZMB(14939)
rat ... GZMB(171528)

General description

GZMB codes for granzyme B.[1]
Granzyme B (gzm B) is a peptidase that induces cytotoxic T lymphocyte (CTL)-mediated DNA fragmentation and apoptosis in allogeneic target cells.[2] It cleaves after aspartic acid residues in substrates. Granzyme B-mediated release of mitochondrial cell death factors are required for the activation of caspase-3. Furthermore, the degradation of granzyme B is known to decrease the susceptibility of cancer cells to NK-cell-mediated lysis during hypoxia. Granzyme B activity assays can be used for studying T cell immunity, CTL responses, cell-mediated cytotoxicity, and apoptosis.[3]
Gzm B is a pro-apoptotic member of the granzyme family.[4] It is a neutral serine protease with Aspase activity. Gzm B is expressed exclusively in the granules of activated cytolytic T cells, natural killer cells and lymphokine-activated killer cells.[2]

Application


  • A Microfluidic Platform for Single Cell Fluorometric Granzyme B Profiling: Describes a technological advancement where the Granzyme B Activity Assay Kit is integrated into a microfluidic platform, enabling high-throughput and precise measurements of Granzyme B activity at the single-cell level, applicable in both research and clinical diagnostics (Briones et al., 2020).

Suitability

Suitable for the detection of granzyme B activity in biological samples and for the characterization of granzyme B inhibitors.

Principle

Granzyme B activity is determined using an enzyme-catalyzed reaction in which the non-fluorescent substrate, Ac-IEPD-AFC, is hydrolyzed resulting in the release of AFC (λex = 380/λem = 500 nm) proportional to the enzymatic activity present. One unit of granzyme B is the amount of enzyme that hydrolyzes 1 pmole of the substrate per minute at 37 °C.

Kit Components Only

Product No.
Description
  • Granzyme B Assay Buffer
  • Granzyme B Substrate
  • Granzyme B Positive Control
  • AFC Standard, 1 mM

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

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Certificates of Analysis (COA)

Lot/Batch Number
6C06K01680

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Granzyme B-induced cell death involves induction of p53 tumor suppressor gene and its activation in tumor target cells.
Meslin F, et al.
The Journal of Biological Chemistry (2007)
Cytotoxic lymphocytes require granzyme B for the rapid induction of DNA fragmentation and apoptosis in allogeneic target cells.
Heusel J W, et al.
Cell, 76(6), 977-987 (1994)
Granzymes: a family of lymphocyte granule serine proteases.
Trapani J A.
Genome Biology, 2(12), 3014-3011 (2001)
Granzyme B?mediated cytochrome c release is regulated by the Bcl-2 family members bid and Bax.
Heibein J A, et al.
The Journal of Experimental Medicine, 192(10), 1391-1402 (2000)
Granzyme B enters the mitochondria in a Sam50-, Tim22-and mtHsp70-dependent manner to induce apoptosis.
Chiusolo V, et al.
Cell Death and Differentiation, 24(4), 747?758-747?758 (2017)
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Questions

1–2 of 2 Questions  

  1. Does the assay buffer in the MAK176 Granzyme B Activity Assay Kit contain protease inhibitors or any substances that help prevent the degradation of the Granzyme B protein?

    1 answer

    1. The assay buffer in MAK176 does not contain any protease inhibitors and granzyme itself is a serine protease.

      Helpful?

  2. Does the Granzyme B Positive Control in the MAK176-1KT Granzyme B Activity Assay Kit require activation before use?

    1 answer

    1. The positive control enzyme does not require activation before use.

      Helpful?

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