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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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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)
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)
Granzymes: a family of lymphocyte granule serine proteases.
Trapani J A.
Genome Biology, 2(12), 3014-3011 (2001)
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)
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)
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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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