Mouse Granzyme B ELISpot Kit Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
PRODUCT SUMMARY
ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. This kit is designed for the detection and enumeration of mouse Granzyme B. Complete ELISpot kits are ready-to-run and require no assay development or refinement.
This ELISpot assay employs a capture antibody specific for mouse Granzyme B, pre-coated onto a PVDF-backed microplate. Appropriately stimulated cells are pipetted directly into the wells and the immobilized antibody in the immediate vicinity of the secreting cells binds secreted mouse Granzyme B. Following wash steps and incubation with a biotinylated detection antibody, alkaline-phosphatase conjugated to streptavidin is added. Unbound enzyme is subsequently removed by washing and a substrate solution (BCIP/NBT) is added. A blue-black colored precipitate forms at the sites of cytokine localization and appears as spots, with each individual spot representing an individual mouse Granzyme B secreting cell. The spots can be counted with an automated ELISpot reader system or manually using a stereomicroscope.
PRODUCT FEATURES
- Detect and quantitate individual cells secreting mouse Granzyme B
- High sensitivity - ELISpot assays can measure responses with frequencies well below 1 in 100,000 cells
- No in vitro expansion of cells required
- High-throughput - ELISpot assays use only a small number of primary cells
KIT CONTENTS
- Mouse Granzyme B Microplate
- Biotinylated Detection Antibody
- Streptavidin conjugated to Alkaline Phosphatase
- Dilution Buffers
- Wash Buffer Concentrate
- BCIP/NBT Chromogen
- Mouse Granzyme B Positive Control
OTHER REAGENTS REQUIRED
- Pipettes and pipette tips
- Deionized or distilled water
- Squirt bottle, manifold dispenser, or automated microplate washer
- 500 mL graduated cylinder
- 37 °C CO2 incubator
- Sterile culture media
- Dissection microscope or an automated ELISpot reader
Product Datasheets
Preparation and Storage
Background: Granzyme B
Granzyme family serine proteases are stored in the granules of cytotoxic T lymphocytes and natural killer cells. Granzymes contain one S1 protease domain and are synthesized as preproproteins. Granzymes are released toward pathogen infected or transformed target cells during cellular immune reactions. Target cell entry by granzymes is through perforin channels. Granzymes trigger apoptosis by caspase-dependent and -independent mechanisms. There are five granzymes (A, B, G, H and K) in human and many more in mouse and rat.
Citations for Mouse Granzyme B ELISpot Kit
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 8
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Glycosite-deleted mRNA of SARS-CoV-2 spike protein as a broad-spectrum vaccine
Authors: CY Wu, CW Cheng, CC Kung, KS Liao, JT Jan, C Ma, CH Wong
Proceedings of the National Academy of Sciences of the United States of America, 2022-03-01;119(9):.
Species: Mouse
Sample Types: Cell Culture Supernates
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LRBA is Essential for Allogeneic Responses in Bone Marrow Transplantation
Sci Rep, 2016-11-08;6(0):36568.
Species: Mouse
Sample Types: Whole Cells
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Suppression of immunodominant antitumor and antiviral CD8+ T cell responses by indoleamine 2,3-dioxygenase.
Authors: Rytelewski, Mateusz, Meilleur, Courtney, Yekta, Maryam A, Szabo, Peter A, Garg, Nitan, Schell, Todd D, Jevnikar, Anthony, Sharif, Shayan, Singh, Bhagirat, Haeryfar, S M Mans
PLoS ONE, 2014-02-28;9(2):e90439.
Species: Mouse
Sample Types: Whole Cells
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Adaptive immune responses elicited by baculovirus and impacts on subsequent transgene expression in vivo.
Authors: Luo, Wen-Yi, Lin, Shih-Yeh, Lo, Kai-Wei, Lu, Chia-Hsi, Hung, Chang-Li, Chen, Chi-Yuan, Chang, Chien-Ch, Hu, Yu-Chen
J Virol, 2013-02-13;87(9):4965-73.
Species: Mouse
Sample Types: Whole Cells
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Radiation-induced loss of cell surface CD47 enhances immune-mediated clearance of human papillomavirus-positive cancer.
Authors: Vermeer D, Spanos W, Vermeer P, Bruns A, Lee K, Lee J
Int J Cancer, 2013-02-12;133(1):120-9.
Species: Mouse
Sample Types: Cell Culture Supernates
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ELISPOT analysis of a new CTL based DNA vaccine for HIV-1 using GM-CSF in DNA prime/peptide boost strategy: GM-CSF induced long-lived memory responses.
Authors: Mahdavi M, Ebtekar M, Khorram Khorshid HR, Azadmanesh K, Hartoonian C, Hassan ZM
Immunol. Lett., 2011-05-23;140(1):14-20.
Species: Mouse
Sample Types: Whole Cells
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Cure of established GL261 mouse gliomas after combined immunotherapy with GM-CSF and IFNgamma is mediated by both CD8+ and CD4+ T-cells.
Authors: Smith KE, Fritzell S, Badn W, Eberstal S, Janelidze S, Visse E, Darabi A, Siesjo P
Int. J. Cancer, 2009-02-01;124(3):630-7.
Species: Mouse
Sample Types: Whole Cells
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A combined DNA vaccine enhances protective immunity against Mycobacterium tuberculosis and Brucella abortus in the presence of an IL-12 expression vector.
Authors: Yu DH, Li M, Hu XD, Cai H
Vaccine, 2007-07-20;25(37):6744-54.
Species: Mouse
Sample Types: Whole Cells
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