Mouse Granzyme B ELISpot Development Module, 5 Plate

Catalog # Availability Size / Price Qty
SEL1865
R&D Systems ELISpot Development Modules
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Citations (7)
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Mouse Granzyme B ELISpot Development Module, 5 Plate Summary

Assay Type
Quantitative Sandwich ELISA Development Module
Format
96-well microplate*
Assay Length
3 hours 35 mins to 4 hours 50 mins**
Sample Type
Whole Cells
Sufficient Materials
For five 96-well microplates***
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

* Sold separately (see Other Reagents Required, below)
** Following an overnight incubation with capture antibody, 2 hour blocking step, and cell stimulation.
*** When using the protocol provided.


PRODUCT SUMMARY
Complete ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. Kits are available for detection and enumeration of a single analyte or two analytes simultaneously. Complete ELISpot kits are ready-to-run and require no assay development or refinement. ELISpot Development Modules contain the basic components required to develop an ELISpot assay. They offer an economical alternative to buying separate antibodies.

ELISpot development modules are an alternative to ELISpot kits. A basic understanding of ELISpot assay development is required for the successful use of these reagents. Each investigator should optimize the coating conditions, the assay sensitivity, the type of enzyme and substrate, as well as the concentrations of the capture and detection antibodies to achieve desired results. The analyte-specific ELISpot Development Module and the ELISpot Blue Color Module contain the necessary components for analyte detection and visualization, respectively. These modules can be used together but are sold separately. Each module contains enough reagents for at least five 96-well microplates.


PRODUCT FEATURES

  • An economical alternative to ELISpot Kits
  • Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
  • Generic development protocols provide direction to start an optimization protocol
  • Customize the assay to your specific needs


MODULE CONTENTS

  • Mouse Granzyme B Capture Antibody
  • Mouse Granzyme B Biotinylated-conjugated Detection Antibody


OTHER REAGENTS REQUIRED

  • ELISpot Blue Color Module or equivalent (R&D Systems, Catalog # SEL002)
  • PBS - 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
  • Wash Buffer - 0.05% Tween® 20 in PBS
  • Blocking Buffer - 1% BSA, 5% Sucrose in PBS
  • Reagent Diluent - 1% BSA in PBS, pH 7.2 - 7.4, 0.2 µm filtered
  • 2 °C – 8 °C refrigerator
  • 37 °C CO2 incubator
  • Positive Control - Use Recombinant Mouse Granzyme B or cells known to secrete Mouse Granzyme B
  • 96-well plates - Nitrocellulose-bottom plates, PVDF-bottom Immunospot® plates, or flat-bottom polystyrene Immulon® ELISA plates
  • Multi-channel pipette, squirt bottle, manifold dispenser, or automated microplate washer
  • Dissection microscope or an automated ELISpot Reader
  • Deionized H2O

Trademarks and registered trademarks are the property of their respective companies.

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

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.

Entrez Gene IDs:
3002 (Human); 14939 (Mouse)
Alternate Names:
C11; CCPI; CGL1; CGL-1; CSPB; CSP-B; CTLA1; CTLA-1; CTSGL1; Fragmentin-2; Granzyme B; Granzyme-2; GRB; GrzB; GZMB; HLP; SECT

Citations for Mouse Granzyme B ELISpot Development Module, 5 Plate

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.

7 Citations: Showing 1 - 7
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  1. Immunological responses following administration of a genotype 1a/1b/2/3a quadrivalent HCV VLP vaccine
    Authors: D Christians, L Earnest-Si, B Chua, P Meuleman, I Boo, B Grubor-Bau, DC Jackson, ZY Keck, SKH Foung, HE Drummer, EJ Gowans, J Torresi
    Sci Rep, 2018;8(1):6483.
    Species: Mouse
    Sample Types: Whole Cells
  2. TIM-3: a novel regulatory molecule of alloimmune activation.
    Authors: Boenisch O, D'Addio F, Watanabe T, Elyaman W, Magee CN, Yeung MY, Padera RF, Rodig SJ, Murayama T, Tanaka K, Yuan X, Ueno T, Jurisch A, Mfarrej B, Akiba H, Yagita H, Najafian N
    J. Immunol., 2010;185(10):5806-19.
    Species: Mouse
    Sample Types: Whole Cells
  3. Oral immunization with a Lactobacillus casei vaccine expressing human papillomavirus (HPV) type 16 E7 is an effective strategy to induce mucosal cytotoxic lymphocytes against HPV16 E7.
    Authors: Adachi K, Kawana K, Yokoyama T, Fujii T, Tomio A, Miura S, Tomio K, Kojima S, Oda K, Sewaki T, Yasugi T, Kozuma S, Taketani Y
    Vaccine, 2010;28(16):2810-7.
    Species: Mouse
    Sample Types: Whole Cells
  4. Enhancing DNA vaccination by sequential injection of lymph nodes with plasmid vectors and peptides.
    Authors: Smith KA, Tam VL, Wong RM, Pagarigan RR, Meisenburg BL, Joea DK, Liu X, Sanders C, Diamond D, Kundig TM, Qiu Z, Bot A
    Vaccine, 2009;27(19):2603-15.
    Species: Mouse
    Sample Types: Whole Cells
  5. IL-12 enhances CTL synapse formation and induces self-reactivity.
    Authors: Markiewicz MA, Wise EL, Buchwald ZS, Cheney EE, Hansen TH, Suri A, Cemerski S, Allen PM, Shaw AS
    J. Immunol., 2009;182(3):1351-61.
    Species: Mouse
    Sample Types: Whole Cells
  6. Mechanisms of immunization against cancer using chimeric antigens.
    Authors: Engelhorn ME, Guevara-Patino JA, Merghoub T, Liu C, Ferrone CR, Rizzuto GA, Cymerman DH, Posnett DN, Houghton AN, Wolchok JD
    Mol. Ther., 2008;16(4):773-81.
    Species: Mouse
    Sample Types: Whole Cells
  7. Tolerization of tumor-specific T cells despite efficient initial priming in a primary murine model of prostate cancer.
    Authors: Anderson MJ, Shafer-Weaver K, Greenberg NM, Hurwitz AA
    J. Immunol., 2007;178(3):1268-76.
    Species: Mouse
    Sample Types: Whole Cells

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