AB46617

Human Granzyme B ELISPOT Kit (with un-coated plates)

Name: Human Granzyme B ELISPOT Kit (with un-coated plates) (ab46617) | Abcam
Brand: Abcam Limited
SKU: AB46617
Price: 220000 JPY
Availability: InStock

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(1 Publication)

Human Granzyme B ELISPOT Kit (with un-coated plates) is a ELISPOT kit for the sensitive detection of Human Granzyme B (with un-coated plates) production in a single cell suspension.

別名を表示する

CGL1, CSPB, CTLA1, GRB, GZMB, Granzyme B, C11, CTLA-1, Cathepsin G-like 1, Cytotoxic T-lymphocyte proteinase 2, Fragmentin-2, Granzyme-2, Human lymphocyte protein, SECT, T-cell serine protease 1-3E, CTSGL1, Lymphocyte protease, HLP

1 Images

ELISPOT - Human Granzyme B ELISPOT Kit (with un-coated plates) (AB46617)

ELISPOT

PubMed

ELISPOT - Human Granzyme B ELISPOT Kit (with un-coated plates) (AB46617)

Measurement of Granzyme B release from Human NK cells using ab46617 - Granzyme B Human Elispot Kit.

NK cells were cultured with rIL-2 and with/without TKD peptide (2 µg/ml) for 4–5 days prior to stimulation with i/uRBC (1 : 3 or 10 : 1), before being isolated. Granzyme B release was determined via ELISPOT assay following the protocol, with 2000 effector cells. Experiments were repeated using a blocking antibody directed against Hsp70.

Image from Böttger E et al., PLoS One. 2012;7(3) : e33774. doi : 10.1371/journal.pone.0033774. Epub 2012 Mar 15.; Fig 6.; March 15, 2012, PLoS ONE 7(3) : e33774.

Böttger E et al., PLoS One, 7, e33774, 2012 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

Key facts

検出方法

Colorimetric

サンプルタイプ

Suspension cells

交差種

Human

アッセイタイプ

Sandwich (qualitative)

アッセイプラットフォーム

Microplate

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ELISPOT": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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製品の詳細

This Human Granzyme B ELISPOT Kit (with un-coated plates) is designed to enumerate Granzyme B producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing Granzyme B production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of Granzuyme B producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. ELISPOT assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.

The ELISPOT assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.

Principle of Method

After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.

Recognizes natural human Granzyme B.

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製品内容

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出荷温度及び保存条件

出荷温度

Blue Ice

短期保存温度

+4°C

長期保存温度

+4°C

保管に関する情報

+4°C

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Granzyme B also known as GZMB GB11 or granzyme B protein is a serine protease with a molecular mass of approximately 32 kDa. It is expressed mainly in cytotoxic T lymphocytes and natural killer (NK) cells. This enzyme plays a mechanical role in inducing apoptosis in target cells serving as an effector protein in the immune system's defense against virally infected cells or transformed cancer cells. The activity of granzyme B relies on its ability to cleave after aspartate residues in substrate proteins leading to the activation of apoptotic pathways.

Biological function summary

Granzyme B participates prominently in the immune response by activating caspases particularly caspase-3 which promotes the breakdown of cellular components necessary for apoptosis. Granzyme B does not function in isolation but acts in concert with other immune system factors such as perforin to effectively induce cell death. Perforin creates pores in the target cell membrane allowing granzyme B to enter and instigate the apoptosis sequence. The enzyme also contributes to the processing of cytokines which enhances the immune response further.

Pathways

Studies have determined that granzyme B is critical in the apoptosis pathway particularly in the granule exocytosis pathway. It closely interacts with proteins such as perforin and other granzymes to mediate apoptosis in target cells. Granzyme B also plays a role in the inflammatory response and can influence pathways associated with cytotoxic T cell signaling. Its pathway interactions ensure effective elimination of damaged or infected cells maintaining tissue homeostasis.

Granzyme B has associations with autoimmune diseases and cancer. Abnormally high levels of granzyme B can contribute to tissue damage and inflammation in autoimmune conditions like rheumatoid arthritis. In the context of cancer granzyme B aids in tumor surveillance and destruction when functioning correctly but impaired granzyme B activity can lead to evasion of immune detection by cancerous cells. Perforin also plays a role in these conditions closely working with granzyme B to either protect against or drive disease progression.

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製品プロトコール

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ターゲットの情報

Abundant protease in the cytosolic granules of cytotoxic T-cells and NK-cells which activates caspase-independent pyroptosis when delivered into the target cell through the immunological synapse (PubMed : 1985927, PubMed : 3262682, PubMed : 3263427). It cleaves after Asp (PubMed : 1985927, PubMed : 8258716). Once delivered into the target cell, acts by catalyzing cleavage of gasdermin-E (GSDME), releasing the pore-forming moiety of GSDME, thereby triggering pyroptosis and target cell death (PubMed : 31953257, PubMed : 32188940). Seems to be linked to an activation cascade of caspases (aspartate-specific cysteine proteases) responsible for apoptosis execution. Cleaves caspase-3, -9 and -10 (CASP3, CASP9 and CASP10, respectively) to give rise to active enzymes mediating apoptosis (PubMed : 9852092). Cleaves and activates CASP7 in response to bacterial infection, promoting plasma membrane repair (By similarity).

See full target information GZMB

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文献 (1)

Recent publications for all applications. Explore the full list and refine your search

PloS one 7:e33774 PubMed22438997

2012

Plasmodium falciparum-infected erythrocytes induce granzyme B by NK cells through expression of host-Hsp70.

Applications

ELISPOT

Species

Human

Evelyn Böttger,Gabriele Multhoff,Jürgen F J Kun,Meral Esen

View all publications

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websiteProtocolBooklet

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Human Granzyme B ELISPOT Kit (with un-coated plates)