Human Granzyme B ELISPOT Kit (with un-coated plates) (ab46617)
Key features and details
- Assay type: Sandwich (qualitative)
- Detection method: Colorimetric
- Sample type: Suspension cells
- Reacts with: Human
製品の概要
-
製品名
Human Granzyme B ELISPOT Kit (with un-coated plates)
Granzyme B キット 製品一覧 -
検出方法
Colorimetric -
サンプルの種類
Suspension cells -
アッセイタイプ
Sandwich (qualitative) -
ステップ
Multiple steps standard assay -
種交差性
交差種: Human -
製品の概要
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.
-
アプリケーション
適用あり: ELISpotmore details -
試験プラットフォーム
Microplate
製品の特性
-
保存方法
Store at +4°C. Please refer to protocols. -
内容 5 x 96 tests 10 x 96 tests 96 PVDF-bottomed-well plates. 5 units 10 units Bovine Serum Albumin 1 x 1g 2 x 1g Dry Skimmed milk 1 x 1g 2 x 1g Biotinylated detection antibody 1 vial 2 vials Granzyme B Capture Antibody 1 x 500µl 2 x 500µl Ready-to-use BCIP/NBT substrate buffer 2 x 27ml 4 x 27ml Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl 2 x 50µl -
研究分野
-
関連性
Cytolytic T lymphocytes and natural killer cells share the remarkable ability to recognize, bind, and lyse specific target cells. They are thought to protect their host by lysing cells bearing on their surface 'nonself' antigens, usually peptides or proteins resulting from infection by intracellular pathogens. Granzyme B is crucial for the rapid induction of target cell apoptosis by CTL in cell-mediated immune response. -
細胞内局在
Cytoplasmic granule. Cytoplasmic granules of cytolytic T-lymphocytes and natural killer cells -
別名
- C11
- Cathepsin G like 1
- CCPI
see all -
参照データベース
- Entrez Gene: 3002 Human
- Omim: 123910 Human
- SwissProt: P10144 Human
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab46617の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| ELISpot |
Use at an assay dependent dilution.
|
| 特記事項 |
|---|
|
ELISpot
Use at an assay dependent dilution. |
画像
-
ELISPOT - Granzyme B Human Elispot Kit (ab46617)Böttger E et al., PLoS One, 7, e33774, 2012 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (1)
ab46617 は 1 報の論文で使用されています。
- Böttger E et al. Plasmodium falciparum-infected erythrocytes induce granzyme B by NK cells through expression of host-Hsp70. PLoS One 7:e33774 (2012). ELISpot ; Human . PubMed: 22438997