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FluoroSpot MAX™ Deluxe Set Human Granzyme B

Pricing & Availability
Regulatory Status
RUO
Other Names
Granzyme-2, GranzymeB, GRB, GrzB, GZMB
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Product Citations
publications
FluoroSpot_Human-Granzyme-B_ELISPOT_092625
Human PBMC (10,000 cells / well) were incubated for 16 hours in the absence or presence of either 1X Cell Activation Cocktail (without Brefeldin A) (Cat. No. 423301) or 1X Cell Activation Cocktail (with Brefeldin A) (Cat. No. 423303). BioLegend’s Cell Activaton Cocktail contains PMA and ionomycin at the optimized concentration. Brefeldin A functions to inhibit intracellular protein transport, as a result, it inhibits secretion of cytokines such as Granzyme B. The number of cells secreting Human Granzyme B was analyzed by FluoroSpot and quantified using an image cytometer equipped with the required fluorescence filter.
  • FluoroSpot_Human-Granzyme-B_ELISPOT_092625
    Human PBMC (10,000 cells / well) were incubated for 16 hours in the absence or presence of either 1X Cell Activation Cocktail (without Brefeldin A) (Cat. No. 423301) or 1X Cell Activation Cocktail (with Brefeldin A) (Cat. No. 423303). BioLegend’s Cell Activaton Cocktail contains PMA and ionomycin at the optimized concentration. Brefeldin A functions to inhibit intracellular protein transport, as a result, it inhibits secretion of cytokines such as Granzyme B. The number of cells secreting Human Granzyme B was analyzed by FluoroSpot and quantified using an image cytometer equipped with the required fluorescence filter.
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439226 5 plates 616€
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Description

Granzyme B is a member of the granzyme family of serine proteases found specifically in granules of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. Perforin/granzyme pathway is a molecular weapon for the defense against virus-infected and malignantly transformed host cells. Perforin protein creates a pore in the membrane of targets cells and facilitate the entry of granzymes. This results in the translocation of Granzyme B into the nucleus and initiation of apoptosis. Other granzymes have also been implicated in apoptosis, but Granzyme B appears to be the most potent. Human Granzyme B is a potential immunoconjugate and an important facilitator in chronic inflammation and wound healing.

BioLegend’s FluoroSpot MAX™ Deluxe Set contains pre-optimized essential reagents, buffers, and solutions for the detection of human Granzyme B secreting cells. In this assay, anti-human Granzyme B capture antibodies are coated on a clear-bottom microplate. Cells (stimulated or unstimulated) are added to the wells and incubated, allowing secreted Granzyme B to bind to nearby capture antibodies. After incubation, cells are removed, and biotinylated detection antibodies are added followed by Streptavidin-PE. The resulting red spots represent the “footprint” of an individual Granzyme B-secreting cell, allowing for precise quantification of cytokine-producing cells in the sample.

Product Details
Technical Data Sheet (pdf)

Kit Details

Kit Contents
  • Human Granzyme B Capture Antibody (200X) 
  • Human Granzyme B Detection Antibody (200X) 
  • Human Granzyme B Standard (used as positive control) 
  • FluoroSpot SA-PE (1000X) 
  • Coating Buffer A (5X) 
  • Assay Diluent B (5X)
Materials Not Included
  • Corning® Clear Bottom Black Polystyrene Microplates, 96 Well (Catalog No. 3601) is recommended, or equivalent black walled, clear bottom microplate is suggested.
  • Black Microplate Lid: Corning® Universal Black Microplate Lid with Corner Notch (Catalog No. 3935) is recommended, or equivalent
  • Wash Buffer: BioLegend Cat. No. 421601 is recommended, or PBS + 0.05% Tween-20  
  • Plate Sealers: BioLegend Cat. No. 423601 is recommended 
  • 0.22 μm filters
  • Sterile PBS (Phosphate-Buffered Saline): 8.0 g NaCl, 1.16 g Na2HPO4, 0.2 g KH2PO4, 0.2 g KCl, add deionized water to 1 L; pH to 7.4, 0.22 μm filtered
  • Sterile cell culture media
  • 37oC CO2 incubator
  • Deionized (DI) water 
  • Adjustable pipettes to measure volumes ranging from 2 μL to 1 mL 
  • Wash bottle, manifold dispenser, or automated microplate washer 
  • Tubes for dilution
  • Timer
  • Alcohol Wipes 
  • Image cytometer or other imaging platform capable of whole-well imaging. The device should be equipped with the appropriate filters for the fluorophores utilized in each assay.

Product Details

Verified Reactivity
Human
Application

FluoroSpot

Antigen Details

Cell Type
Basophils, Dendritic cells, Leukocytes, Lymphocytes, Macrophages, Mast cells, Monocytes, NK cells, NKT cells, T cells, Th1, Tregs
Biology Area
Adaptive Immunity, Apoptosis/Tumor Suppressors/Cell Death, Cancer Biomarkers, Cell Death, Immuno-Oncology, Immunology, Innate Immunity, Mitochondrial Function, Neuroinflammation, Signal Transduction, Ubiquitin/Protein Degradation
Molecular Family
CD Molecules, Cytokine/Chemokine Receptors, Cytokines/Chemokines, Enzymes and Regulators, Immune Checkpoint Receptors, Innate Immune Signaling, MHC Antigens, Mitochondrial Markers, Proteases, TCRs, Toll Like Receptors, Tumor Suppressors
Gene ID
3002 View all products for this Gene ID
UniProt
View information about Granzyme B on UniProt.org

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Go To Top Version: 1    Revision Date: 09/26/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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