Halo^® PMT

A Standardized and Validated Bioluminescence Assay to Monitor Lympho-Hematopoietic Reconstitution After Transplantation

Description	Tissue	Formulation	Catalog Number	Quantity
Multiple
4-Population + Background	Bone marrow or peripheral blood	Low serum	K2-4PMT-5	1 Kit
5-Population + Background	Bone marrow or peripheral blood	Low serum	K2-5PMT-6	1 Kit
7-Population + Background	Bone marrow or peripheral blood	Low serum	K2-7PMT-8	1 Kit
Multiple
4-Population + Background	Bone marrow or peripheral blood	Serum-Free	K2SF-4PMT-5	1 Kit
5-Population + Background	Bone marrow or peripheral blood	Serum-Free	K2SF-5PMT-6	1 Kit
7-Population + Background	Bone marrow or peripheral blood	Serum-Free	K2SF-7PMT-8	1 Kit

 

• Determine short- and long-term reconstitution of stem cells and multiple lineages, include those of the immune system (T- and B-cell lineages).
• Short 5-7 day incubation period allows reconstitution to be monitored continually.
• Quantitative cell population proliferation data provides all the information to predict and monitor lympho-hematopoietic reconstitution.
• Suspension Expansion Culture™ (SEC™) Technology provides the accuracy and precision needed and makes the use of any methylcellulose CFU assay obsolete.
• Incorporates ATP Bioluminescence Technology; the most sensitive and accurate readout available.
• Fully standardized, verfied and validated according to FDA guidelines.
• Proficiency testing performed and completed during the assay standardization procedure. No additional costly and time-consuming testing required.
• Measurement assurance parameters provide proficiency and the trust in your results.
• 96-well plate format ensures smaller sample and reagent volumes with faster setup.
• Multiplexes with flow cytometry and other readouts.
• Let the plate reader acquire and calculate results in just 5 minutes or less.
• Directly compare reconstitution of different tissues, donors and patients over time to obtain important historical data for later use.
• Includes everything needed to culture and measure 4-, 5- or 7-populations for "global" assessment of multi-lineage hematopoietic or multi-llineage ympho-hematopoietic reconstitution.
• Time-efficient and cost-effect.
• Easy to learn in just 1 day.

 

All ATP bioluminescence assays kits from Preferred Cell Systems™ include an ATP standard(s) and controls.  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.


 

• 4-Population "Global" Assay consists of: SC-GEMM 1, P-BFU 1, P-GM 1, P-Mk 1 and Background
• 5-Population "Global Assay consists of: SC-HPP 2, SC-GEMM 1, P-BFU 1, P-GM 1, P-Mk 1 and Background
• 7-Population "Global" Assay consists of: SC-HPP 2, SC-GEMM 1, P-BFU 1, P-GM 1, P-Mk 1, P-Tcell 3, P-Bcell 2 and Background

The 4-, 5- and 7-populations assays contain 5, 6 and 8 x 96-well plates, one for each lineage and background. HALO® PMT "Global" is meant to be performed using 8 replicates/sample, but this can be reduced to 4 replicates/sample. Using 8 replicates/sample, the maximum number of samples that can be assessed/plate is 12.

• Umbilical cord blood
• Peripheral blood
• Bone marrow biopsy

Tissues used for Detection

• Peripheral blood
• Bone marrow biopsy

The recommended cell purity is a mononuclear cell (MNC) fraction. A total nucleated cell (TNC) fraction is not recommended as this contains high concentrations of cell impurities, such as red blood cells, neutrophils, platelets and other cells that severely underestimate and even inhibit the detection of progenitor cells. 

For Research Use Only. Not for clinical diagnostic use.

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

• HALO® PMT "Global" Master Mixes
• ATP standard
• ATP high and low controls
• ATP Enumeration Reagent
• Sterile, 96-well plates
• Non-sterile, 96-well plates
• Sterile, adhesive foil covers

A specific video tutorial on using the PMT assays is not yet ready. Please scroll down and check out the QuickGuide and Technical Manual (if available). Below, are the links to perform the ATP bioluminescence readout:

How to Calibrate and Standardize an ATP Bioluminescence Assay - Part 1 (for HALO® PMT)

How to Calibrate and Standardize an ATP Bioluminescence Assay - Part 2 (for HALO® PMT)

Proficiency Testing for Hematopoietic Cellular Therapy Products
 

Download the HALO® PMT "Global" Technical Manual
Download the ATP Optimization Kit Protocol for First-Time Users
Download the Luminometer Setup and RLU to ATP Conversion Procedure
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