ImmunoGLO™, ImmunoGLO™ TCP,
ImmunoGLO™ BCP, ImmunoGLO™ MLC

A Family of In Vitro Bioluminescence Assays for
Immune Cell Proliferation, T- and B-Cells and
Mixed Lymphocyte Culture

 

 

Description Catalog Number Quantity
An Add-On Kit for Any Bioluminescence "GLO" Assay K-ATPSC-1 1 Kit

ImmunoGlo™

An In Vitro Bioluminescence Basic Viability and Proliferation Assay for Immune Cells

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
Immune Cells Multiple / Any ImmunoGro™ Base Medium KM1-LPA-1 1 Kit

ImmunoGlo™ TCP

An In Vitro Bioluminescence T-Lymphocyte Proliferation Assay

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
T-lymphocytes Human / Any Low Serum + IL-2 KM1-TCP1-1H 1 Kit
T-lymphocytes Human / Any Low Serum + CD3 + CD28 KM1-TCP2-1H 1 Kit
T-lymphocytes Human / Any Low Serum + IL-2 + CD3 + CD28 KM1-TCP3-1H 1 Kit
T-lymphocytes Human / Any Serum-free + IL-2 KM1SF-TCP1-1H 1 Kit
T-lymphocytes Human / Any Serum-free + CD3 + CD28 KM1SF-TCP2-1H 1 Kit
T-lymphocytes Human / Any Serum-free + IL-2 + CD3 + CD28 KM1SF-TCP3-1H 1 Kit

ImmunoGlo™ TCP

An In Vitro Bioluminescence T-Lymphocyte Proliferation Assay

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
T-lymphocytes Primate / Any Low Serum + IL-2 KM1-TCP1-1Pr 1 Kit

ImmunoGlo™ TCP

An In Vitro Bioluminescence T-Lymphocyte Proliferation Assay

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
T-lymphocytes Rat / Any Low Serum + IL-2 KM1-TCP1-1R 1 Kit

ImmunoGlo™ TCP

An In Vitro Bioluminescence T-Lymphocyte Proliferation Assay

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
T-lymphocytes Mouse / Any Low Serum + IL-2 KM1-TCP1-1M 1 Kit

ImmunoGlo™ BCP

An In Vitro Bioluminescence T-Lymphocyte Proliferation Assay

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
B-lymphocytes Human / Any Low Serum +Ligands+IL4 KM1-BCP2-1H 1 Kit
B-lymphocytes Human / Any Low Serum+Conjugates+Ligands+IL-4 KM1-BCP3-1H 1 Kit

ImmunoGlo™ BCP

An In Vitro Bioluminescence T-Lymphocyte Proliferation Assay

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
B-lymphocytes Mouse / Any Low Serum+Conjugates+IL-4 KM1-BCP3-1M 1 Kit

ImmunoGlo™ MLC

An In Vitro Bioluminescence 1- or 2-Way Mixed Lymphocyte Culture (MLC) Assay

Cell Population Species/Tissue ImmunoGro™ Medium Formulation Catalog Number Quantity
Immune Cells Multiple / Any Includes Mitomycin-C for 1-way MLC KM1-MLC-1 1 Kit

 

ImmunoGLO™: A family of immune proliferation assays for T- and B-lymphocytes and mixed lymphocyte culture/reaction (MLC/MLR) assays

 

 

  • ImmunoGlo™ is a general use functional immune cell proliferation assay where the investigator employs their own culture reagents and protocol for any type of immune cell.
  • ImmunoGlo™ TCP Assay Kits include specific cocktails to stimulate functional T-lymphocytes.
  • ImmunoGlo™ BCP is specific for stimulating functional human B-lymphocytes.
  • ImmunoGlo™ MLC for a 1- or 2-way mixed lymphocyte culture (MLC) or reaction (MLR).
All assays incorporate reagents to perform a standardized ATP bioluminescence readout. 
 
Uses of the ImmunoGlo™ family of assays include, but are not limited to, the following:
 
  • Lymphocyte activation and proliferation.
  • Test unprimed T-cells in the presence of antibodies, enterotoxins, mitogens etc.
  • CAR-T cell studies.
  • Cellular immune response studies.
  • Single-cell, lymphocyte cloning studies.
  • Test DLI (Donor Lymphocyte Infusion) samples for stimulation/induction prior to use.
  • Effect of accessory lymphocyte induction.
  • Effect of co-stimulators on lymphocyte induction.
  • Effect of epitope sequences and novel peptides or proteins.
  • Test the response of primed T-cells in vitro.
  • In vitro immunotoxicity studies (see Immuno™-Tox HT).
  • Immune cell gene targeting and expression, e.g. CRISPR-Cas9.
Uses of ImmunoGlo™ MLC include:
  • One-way or two-way mixed lymphocyte culture (MLC) / reaction (MLR).
  • Allogeneic mixed lymphocyte culture.
  • Dendritic cell interactions.
  • Mesenchymal stem/stromal cell interactions.
  • Agents that activate/suppress MLC/MLR.
 
The ImmunoGlo™ family of assays are the most advanced assays to measure immune cell response. The most important benefits include:  
  • The most sensitive, accurate and reliable, non-radioactive assay incorporating proven ATP bioluminescence technology.
  • Instrument-based, quantitative and fully standardized allowing comparison of results over time.
  • 10 - 100 times more sensitive than WST-1, CSFE and other absorbance or fluorescence assays.
  • High throughput capability.
  • Ability to use any mitogens, co-stimulators etc and lymphocyte culture protocols (ImmunoGlo™).
  • Ability to stimulate T-cells using IL-2 alone, CD3 and CD28 or IL-2, CD3 and CD28 cocktails ImmunoGlo™ TCP).
  • Ability to specifically study naive or activated B-lymphocyte populations (ImmunoGlo™ BCP).
  • Multiplexing capability to incorporate flow cytometry, cytokine release and other assay readouts for additional information from the same sample.
  • Optional ATP Bioluminescence Standardization & Calibration Kit (Cat. No. K-ATPSC-1) available (see below).

Principle of ATP bioluminescence procedure for all ImmunoGlo™ assays

If required, ImmunoGlo™, ImmunoGlo™ TCP and ImmunoGlo™ BCP can be optionally standardized using the ATP Bioluminescence Standardization and Calibration Kit (Cat. No. K-ATPSC-1). ImmunoGlo™ MLC includes all reagents and supplies to standardize the assay.  Performing the ATP standard curve and controls is highly recommended. The controls are used to calibrate the luminescence plate reader and the ATP standard is used to standardize the assay. The values obtained from the standard curve and controls (see diagram below) are then compared to the measurement assurance parameters that are provided in the instruction manual. If the values obtained are within the ranges provided by the measurement assurance parameters, you, the user, have successfully performed the proficiency testing to ensure that you have not only performed the assay correctly, but that the results you obtain can be trusted.

No additional proficiency testing is required if the calibration and standardization procedure is performed. The values you obtain from each calibration and standardization can be logged and used for certification that the assay has been performed correctly and that the results are trustworthy.

Calibration and standardization of an ATP bioluminescence assay from Preferred Cell Systems™ - Measurement Assurance
 
ImmunoGlo™
  • Multiple
ImmunoGlo™ TCP
  • Multiple, but not all assays available for all species. Please see online catalog.
ImmunoGlo™ BCP
  • Human
  • Mouse
  • Peripheral blood mononuclear cells (PBMC)
  • Umbilical cord blood
  • Bone marrow
  • Lymphocyte sub-populations
  • Lymphocyte tissues and organs (e.g. spleen, lymph nodes)
The diagram below show how T-cells can be stimulated and detected using the ImmunoGlo™ TCP Assay Kit
Detection of T-Cell Stimulation using ImmunoGlo™ TCP

The diagrams below show an example of a 1-way and a 2-way mixed lymphocyte culture (reaction) using ImmunoGlo™ MLC assay kits.
1-Way Mixed Lymphocyte Culture (Reaction) using ImmunoGlo™ MLC

2-Way Mixed Lymphocyte Culture (Reaction) using ImmunoGlo™ MLC

 

For Research Use Only. Not for clinical diagnostic use.

Luminescence or multimode plate reader with "glow" luminescence measuring capability.

All ImmunoGlo™, ImmunoGlo™ TCP and ImmunoGlo™ BCP contain:
  • ImmunoGro™ Medium
  • ATP Enumeration Reagent
  • Sterile, 96-well culture plates
  • Sterile, adhesive plate cover foils to maintain sterility of unused wells
PLEASE NOTE: If required, all ImmunoGlo™, ImmunoGlo™ TCP and ImmunoGlo™ BCP assays can be standardized using the add-on ATP Bioluminescence Standardization and Calibration Kit, Catalog Number K-ATPSC-1. ImmunoGlo™ MLC includes calibration and standardization reagents and supplies.

ImmunoGlo™ TCP assay kits are available with the choice of:
  • IL-2 alone, or
  • CD3 and CD28 co-stimulators, or
  • IL-2 + CD3 + CD28 co-stimulators as a complete cocktail
ImmunoGlo™ BCP assay kits for human or mouse B-cells are available with the following cocktails:
  • Proprietary cocktail contains cytokines only.
  • Proprietary cocktail includes conjugates, ligands and cytokines (human)
  • Proprietary cocktail includes conjugates and cytokines (mouse).
ImmunoGlo™ MLC contains the same contents for all ImmunoGlo™ assays, but includes:
  • Mitomycin-C to perform a 1-way MLC
  • IMDM for serial dilution of the ATP standard
  • ATP standard
  • ATP controls
  • ATP Enumeration Reagent
  • Sterile, 96-well culture plates
  • Non-sterile, 96-well plates for ATP standard curve
  • Sterile, adhesive plate cover foils to maintain sterility of unused wells



Complete, bioluminescence functionality and proliferation assay for immune cells.

ImmunoGlo™ TCP assays for T-lymphocytes


ImmunoGlo™ BCP assays for B-lymphocytes

Bioluminescence 1- or 2-Way Mixed Lymphocyte Culture (MLC) Assay

 

Download the ImmunoGlo™ Assay Technical Manual
Download the ImmunoGlo™ TCP Assay Technical Manual
Download the ImmunoGlo™ BCP Assay Technical Manual
Download the ImmunoGlo™ MLC Assay Technical Manual
Download the ATP Bioluminescence Standardization & Calibration Kit Manual
Download Luminometer setup and RLU to ATP conversion (for ImmunoGlo™ MLC)
Download Certificate of Analysis (CoAs) for ATP Enumeration Reagent (ATP-ER)
Download Certificate of Analysis for ATP Stanadrds
Download Certificate of Analysis for ATP Controls
Download Certificate of Analysis of ATP Reconstitution Reagent
Download Certificate of Analysis for Sterile 96-Well Plates
Download Certificate of Analysis for Non-Sterile, 96-Well Plates
Download Certificate of Analysis for Sealing Films
Download Certificate of Analysis for IMDM