Mouse CD4+/Granzyme B ELISpot Kit

Catalog # Availability Size / Price Qty
EL6024
Product Details
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Mouse CD4+/Granzyme B ELISpot Kit Summary

Assay Type
Quantitative Sandwich ELISA
Format
96-well PVDF-backed microplate
Assay Length
3 hours 15 mins to 4 hours 30 mins*
Sample Type
Whole Cells
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.

* Following cell stimulation.


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

Stability & Storage
Store the unopened product at 2 - 8 °C. 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

FAQs

No product specific FAQs exist for this product, however you may

View all ELISpot and FluoroSpot Kit FAQs

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Mouse CD4+/Granzyme B ELISpot Kit
By Anonymous on 11/06/2017
Sample Tested: 2120Ep human embryonal carcinoma cell line