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QuickZyme Human Granzyme B Activity Assay Reagent Set 2X 96-Assays

  • Regulatory status:RUO
  • Type:ELISA development kit
  • Other names:Granzyme 2, Fragmentin 2, Human lymphocyte protein, Granzyme-B, GrB, GZMB, C11, CTLA-1, Cathepsin G-like 1, CTSGL1, Cytotoxic T-lymphocyte proteinase 2, Lymphocyte protease, HLP, SECT, T-cell serine protease 1-3E
  • Species:Human, Mouse
Cat. No. Size Price


New QZBGRB2 Reagent set for two 96-well plates $1467,42
PubMed Product Details
Technical Data

Type

ELISA development kit

Description

The QuickZyme Granzyme B Activity Assay Reagent Set (two 96-well plates per set) allows quantitative measurement of active human Granzyme B in a variety of sample types, such as conditioned cell culture media, cell extracts, body fluids (plasma, wound fluid), tissue extracts, and (purified) enzyme preparations. The assay also recognizes mouse Granzyme B. The easy and sensitive immunoassay has the ability to detect both high and low ranges at a low cost per sample.

Applications

Tissue extract, Plasma, Body fluids, Cells/cell extracts, Cell culture conditioned medium, Purified enzyme preparations

Sample Requirements

10 - 100 μl

Shipping

On dry ice. Upon receipt, store the product at the temperature recommended below.

Storage/Expiration

Antibody stock: Store at -20°C, avoid repeated freeze/thaw cycles. Granzyme B standard: Store at -70°C, avoid repeated freeze/thaw cycles. Detection enzyme: Store at -70°C, avoid repeated freeze/thaw cycles. Chromogenic Substrate: Store at -20°C.

Calibration Range

0 - 10 ng/ml (high activity, 2 or 4 h incubation)

0 - 0.31 ng/ml (low activity, 24 h incubation)

Limit of Detection

0.01 ng/ml (24 hr incubation)

Summary

Features

  • Reagent set for two 96-well plates

  • Measures active human Granzyme B

  • Also recognizes mouse Granzyme B (Eur. J. Immunol 2012, 264-266)

  • Samples: conditioned cell culture media, cell extracts, body fluids (plasma, wound fluid), tissue extracts, and (purified) enzyme preparations

  • Quantitative

  • Range/sensitivity: 0.01 ng/ml (24 h incubation)

  • Ease-of-use: Equivalent to ELISA Overnight protocol

  • Reagent set contains antibody, standard, detection enzyme and substrate

Research topic

Apoptosis, Extracellular matrix, Immune Response, Infection and Inflammation, Transplantation

Summary

Granzymes are exogenous serine proteinases (enzymes) that are released from cytoplasmic granules of cytotoxic lymphocytes (CTLs) and NK cells.

The name “granzymes” is derived from: granules + enzymes. These granules contain next to granzymes other proteins including a pore-forming protein (Perforin). Upon binding of the CTL to a target cell (by CTL-receptor and antigen-presenting MHC molecules on the target cell) the contents of the granules are released in the intercellular space where after perforin will “perforate” the target cell membrane by forming transmembrane pores. Through these pores the granzymes can now enter the cytosol of the target cell. Granzyme B activates the intracellular cascade of caspases finally resulting in the killing of the target cells.

Not all granzymes enter the target cell, part of them also “leak” in to the peripheral blood and other biological fluids. Detectable amounts of granzymes have been found to circulate in healthy volunteers. These soluble granzymes can be measured by ELISA.

Granzyme B is also capable of acting extracellularly (independent of perforin) and influences extracellular matrix (ECM) degradation and remodeling by cleavage of several important ECM proteins, including laminin, decorin, fibronectin and vitronectin. Thus, Granzyme B may play an important role in thinning of aging skin, wound healing, and many chronic inflammatory diseases where ECM degradation is a hallmark.

Viral infections: Increased levels of soluble granzymes have been found with patients suspected of an increased NK cell and CTL-response caused by systemic viral infections such as EBV, HIV, CMV, hepatitis A and Dengue fever.

Lymphomas and carcinomas: It is shown that the presence of a high percentage of Granzyme B positive CTLs in glands of patients suffering from Hodgkin’s disease correlate with a severe prognosis.

Rheumatoid arthritis: Soluble Granzyme A and B are increased in synovial fluid from rheumatoid arthritis and significantly higher than levels in patients with osteoarthrosis.

Transplantation: Granzymes are likely involved in the acute rejection of kidney-transplants, as infiltrating lymphocytes in the rejected kidney strongly express granzymes. Increasing plasma levels of soluble granzymes in patients with a kidney transplants suggest a systemic viral infection, in particular an infection by CMV.

Summary References (10)

References to Granzyme B

  • Boivin WA, Cooper DM, Hiebert PR, Granville DJ. Intracellular versus extracellular granzyme B in immunity and disease: challenging the dogma. Lab Invest. 2009 Nov;89(11):1195-220. doi: 10.1038/labinvest.2009.91. Epub 2009 Sep 21. PMID: 19770840; PMCID: PMC7102238. See more on PubMed
  • Buzza MS, Zamurs L, Sun J, Bird CH, Smith AI, Trapani JA, Froelich CJ, Nice EC, Bird PI. Extracellular matrix remodeling by human granzyme B via cleavage of vitronectin, fibronectin, and laminin. J Biol Chem. 2005 Jun 24;280(25):23549-58. doi: 10.1074/jbc.M412001200. Epub 2005 Apr 19. PMID: 15843372. See more on PubMed
  • Hiebert PR, Boivin WA, Abraham T, Pazooki S, Zhao H, Granville DJ. Granzyme B contributes to extracellular matrix remodeling and skin aging in apolipoprotein E knockout mice. Exp Gerontol. 2011 Jun;46(6):489-99. doi: 10.1016/j.exger.2011.02.004. Epub 2011 Feb 18. PMID: 21316440. See more on PubMed
  • Hiebert PR, Wu D, Granville DJ. Granzyme B degrades extracellular matrix and contributes to delayed wound closure in apolipoprotein E knockout mice. Cell Death Differ. 2013 Oct;20(10):1404-14. doi: 10.1038/cdd.2013.96. Epub 2013 Aug 2. PMID: 23912712; PMCID: PMC3770318. See more on PubMed
  • Kurschus FC, Jenne DE. Delivery and therapeutic potential of human granzyme B. Immunol Rev. 2010 May;235(1):159-71. doi: 10.1111/j.0105-2896.2010.00894.x. PMID: 20536562. See more on PubMed
  • Lord SJ, Rajotte RV, Korbutt GS, Bleackley RC. Granzyme B: a natural born killer. Immunol Rev. 2003 Jun;193:31-8. doi: 10.1034/j.1600-065x.2003.00044.x. PMID: 12752668. See more on PubMed
  • Parkinson LG, Toro A, Zhao H, Brown K, Tebbutt SJ, Granville DJ. Granzyme B mediates both direct and indirect cleavage of extracellular matrix in skin after chronic low-dose ultraviolet light irradiation. Aging Cell. 2015 Feb;14(1):67-77. doi: 10.1111/acel.12298. Epub 2014 Dec 11. PMID: 25495009; PMCID: PMC4326907. See more on PubMed
  • Rousalova I, Krepela E. Granzyme B-induced apoptosis in cancer cells and its regulation (review). Int J Oncol. 2010 Dec;37(6):1361-78. doi: 10.3892/ijo_00000788. PMID: 21042704. See more on PubMed
  • Shi L, Yang X, Froelich CJ, Greenberg AH. Purification and use of granzyme B. Methods Enzymol. 2000;322:125-43. doi: 10.1016/s0076-6879(00)22013-2. PMID: 10914010. See more on PubMed
  • Trapani JA, Sutton VR. Granzyme B: pro-apoptotic, antiviral and antitumor functions. Curr Opin Immunol. 2003 Oct;15(5):533-43. doi: 10.1016/s0952-7915(03)00107-9. PMID: 14499262. See more on PubMed
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