HALO™

The In Vitro Standardized and Validated Bioluminescence Research Assay Platform for Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 1	Human	Low serum. Methylcellulose-free	K2-HPP1-1H	1 Kit
SC-HPP 2	Human	Low serum. Methylcellulose-free	K2-HPP2-1H	1 Kit
SC-GEMM 1	Human	Low serum. Methylcellulose-free	K2-GEMM1-1H	1 Kit
SC-GEMM 2	Human	Low serum. Methylcellulose-free	K2-GEMM2-1H	1 Kit
SC-GEMM 3	Human	Low serum. Methylcellulose-free	K2-GEMM3-1H	1 Kit
SC-GEM 1	Human	Low serum. Methylcellulose-free	K2-GEM1-1H	1 Kit
SC-GEM 3	Human	Low serum. Methylcellulose-free	K2-GEM3-1H	1 Kit
SC-GEM 2	Human	Low serum. Methylcellulose-free	K2-GEM2-1H	1 Kit
Stem Cells
SC-HPP1T	Human	Serum-free. Methylcellulose-free	K2SF-HPP1T-1H	1 Kit
SC-HPPX	Human	Serum-free. Methylcellulose-free	K2SF-HPPX-1H	1 Kit
SC-HPPXT	Human	Serum-free. Methylcellulose-free	K2SF-HPPXT-1H	1 Kit
SC-HPP 1	Human	Serum-free. Methylcellulose-free	K2SF-HPP1-1H	1 Kit
SC-HPP 2	Human	Serum-free. Methylcellulose-free	K2SF-HPP2-1H	1 Kit
SC-GEMM 1	Human	Serum-free. Methylcellulose-free	K2SF-GEMM1-1H	1 Kit
SC-GEMM 2	Human	Serum-free. Methylcellulose-free	K2SF-GEMM2-1H	1 Kit
SC-GEMM 3	Human	Serum-free. Methylcellulose-free	K2SF-GEMM3-1H	1 Kit
SC-GEM 1	Human	Serum-free. Methylcellulose-free	K2SF-GEM1-1H	1 Kit
SC-GEM 2	Human	Serum-free. Methylcellulose-free	K2SF-GEM2-1H	1 Kit
SC-GEM 3	Human	Serum-free. Methylcellulose-free	K2SF-GEM3-1H	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Human	Low serum. Methylcellulose-free	K2-BFU1-1H	1 Kit
P-BFU 2	Human	Low serum. Methylcellulose-free	K2-BFU2-1H	1 Kit
P-GM 1	Human	Low serum. Methylcellulose-free	K2-GM1-1H	1 Kit
P-GM 2	Human	Low serum. Methylcellulose-free	K2-GM2-1H	1 Kit
P-GM 3	Human	Low serum. Methylcellulose-free	K2-GM3-1H	1 Kit
P-Mk 1	Human	Low serum. Methylcellulose-free	K2-Mk1-1H	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Human	Serum-free. Methylcellulose-free	K2SF-BFU1-1H	1 Kit
P-BFU 2	Human	Serum-free. Methylcellulose-free	K2SF-BFU2-1H	1 Kit
P-GM 1	Human	Serum-free. Methylcellulose-free	K2SF-GM1-1H	1 Kit
P-GM 2	Human	Serum-free. Methylcellulose-free	K2SF-GM2-1H	1 Kit
P-GM 3	Human	Serum-free. Methylcellulose-free	K2SF-GM3-1H	1 Kit
P-Mk 1	Human	Serum-free. Methylcellulose-free	K2SF-Mk1-1H	1 Kit
Lymphopoietic Progenitor Cells
P-Tcell 1	Human	Low serum. Methylcellulose-free	K2-T1-1H	1 Kit
P-Tcell 2	Human	Low serum. Methylcellulose-free	K2-T2-1H	1 Kit
P-Tcell 3	Human	Low serum. Methylcellulose-free	K2-T3-1H	1 Kit
P-Bcell 2	Human	Low serum. Methylcellulose-free	K2-B2-1H	1 Kit
P-Bcell 3	Human	Low serum. Methylcellulose-free	K2-B3-1H	1 Kit
Lymphopoietic Progenitor Cells
P-Tcell 1	Human	Serum-free. Methylcellulose-free	K2SF-T1-1H	1 Kit
P-Tcell 2	Human	Serum-free. Methylcellulose-free	K2SF-T2-1H	1 Kit
P-Tcell 3	Human	Serum-free. Methylcellulose-free	K2SF-T3-1H	1 Kit
No Growth Factors Added
Any	Human	Low serum. Methylcellulose-free	K2-0-1H	1 Kit
Any	Human	Serum-free. Methylcellulose-free	K2SF-0-1H	1 Kit
"Global" Research Assays
"Global" 4-Pop Research	Human	Low serum, methylcellulose-free	K2-4P-5H	1 Kit
"Global" 5-Pop Research	Human	Low serum, methylcellulose-free	K2-5P-6H	1 Kit
"Global" 7-Pop Research	Human	Low serum, methylcellulose-free	K2-7P-8H	1 Kit
"Global" Research Assays
"Global" 4-Pop Research	Human	Serum-free. Methylcellulose-free	K2SF-4P-5H	1 Kit
"Global" 5-Pop Research	Human	Serum-free. Methylcellulose-free	K2SF-5P-6H	1 Kit
"Global" 7-Pop Research	Human	Serum-free. Methylcellulose-free	K2SF-7P-8H	1 Kit

HALO™

The Easy and Rapid In Vitro Research Bioluminescence Assay for Measuring Cell Viability and Proliferation of Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 2	Mouse	Low serum. Methylcellulose-free	K2-HPP2-1M	1 Kit
SC-GEMM 1	Mouse	Low serum. Methylcellulose-free	K2-GEMM1-1M	1 Kit
Stem Cells
SC-HPP 2	Mouse	Serum-free. Methylcellulose-free	K2SF-HPP2-1M	1 Kit
SC-GEMM 1	Mouse	Serum-free. Methylcellulose-free	K2SF-GEMM1-1M	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Mouse	Low serum. Methylcellulose-free	K2-BFU1-1M	1 Kit
P-BFU 2	Mouse	Low serum. Methylcellulose-free	K2-BFU2-1M	1 Kit
P-GM 1	Mouse	Low serum. Methylcellulose-free	K2-GM1-1M	1 Kit
P-GM 3	Mouse	Low serum. Methylcellulose-free	K2-GM3-1M	1 Kit
P-Mk 1	Mouse	Low serum. Methylcellulose-free	K2-Mk1-1M	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Mouse	Serum-free. Methylcellulose-free	K2SF-BFU1-1M	1 Kit
P-BFU 2	Mouse	Serum-free. Methylcellulose-free	K2SF-BFU2-1M	1 Kit
P-GM 1	Mouse	Serum-free. Methylcellulose-free	K2SF-GM1-1M	1 Kit
P-GM 3	Mouse	Serum-free. Methylcellulose-free	K2SF-GM3-1M	1 Kit
P-Mk 1	Mouse	Serum-free. Methylcellulose-free	K2SF-Mk1-1M	1 Kit
Lymphopoietic Progenitor Cells
P-Tcell 1	Mouse	Low serum. Methylcellulose-free	K2-T1-1M	1 Kit
P-Bcell 3	Mouse	Low serum. Methylcellulose-free	K2-B3-1M	1 Kit
Lymphopoietic Progenitor Cells
P-Tcell 1	Mouse	Serum-free. Methylcellulose-free	K2SF-T1-1M	1 Kit
No Growth Factors Added
Any	Mouse	Low serum. Methylcellulose-free	K2-0-1M	1 Kit
Any	Mouse	Serum-free. Methylcellulose-free	K2SF-0-1M	1 Kit
"Global" Research Assays
"Global" 4-Pop Research	Mouse	Low serum, methylcellulose-free	K2-4P-5M	1 Kit
"Global" 5-Pop Research	Mouse	Low serum, methylcellulose-free	K2-5P-6M	1 Kit
"Global" 7-Pop Research	Mouse	Low serum, methylcellulose-free	K2-7P-8M	1 Kit
"Global" Research Assays
"Global" 4-Pop Research	Mouse	Serum-free. Methylcellulose-free	K2SF-4P-5M	1 Kit
"Global" 5-Pop Research	Mouse	Serum-free. Methylcellulose-free	K2SF-5P-6M	1 Kit
"Global" 7-Pop Research	Mouse	Serum-free. Methylcellulose-free	K2SF-7P-8M	1 Kit

HALO™

The Easy and Rapid In Vitro Research Bioluminescence Assay for Measuring Cell Viability and Proliferation of Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 2	Primate	Low serum. Methylcellulose-free	K2-HPP2-1Pr	1 Kit
SC-GEMM 1	Primate	Low serum. Methylcellulose-free	K2-GEMM1-1Pr	1 Kit
Stem Cells
SC-HPP 2	Primate	Serum-free. Methylcellulose-free	K2SF-HPP2-1Pr	1 Kit
SC-GEMM 1	Primate	Serum-free. Methylcellulose-free	K2SF-GEMM1-1Pr	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Primate	Low serum. Methylcellulose-free	K2-BFU1-1Pr	1 Kit
P-BFU 2	Primate	Low serum. Methylcellulose-free	K2-BFU2-1Pr	1 Kit
P-GM 1	Primate	Low serum. Methylcellulose-free	K2-GM1-1Pr	1 Kit
P-GM 3	Primate	Low serum. Methylcellulose-free	K2-GM3-1Pr	1 Kit
P-Mk 1	Primate	Low serum. Methylcellulose-free	K2-Mk1-1Pr	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Primate	Serum-free. Methylcellulose-free	K2SF-BFU1-1Pr	1 Kit
P-BFU 2	Primate	Serum-free. Methylcellulose-free	K2SF-BFU2-1Pr	1 Kit
P-GM 1	Primate	Serum-free. Methylcellulose-free	K2SF-GM1-1Pr	1 Kit
P-GM 3	Primate	Serum-free. Methylcellulose-free	K2SF-GM3-1Pr	1 Kit
P-Mk 1	Primate	Serum-free. Methylcellulose-free	K2SF-Mk1-1Pr	1 Kit
Lymphopoietic Progenitor Cells
P-Tcell 1	Primate	Low serum. Methylcellulose-free	K2-T1-1Pr	1 Kit
Lymphopoietic Progenitor Cells
P-Tcell 1	Primate	Serum-free. Methylcellulose-free	K2SF-T1-1Pr	1 Kit
No Growth Factors Added
Any	Primate	Low serum. Methylcellulose-free	K2-0-1Pr	1 Kit
Any	Primate	Serum-free. Methylcellulose-free	K2SF-0-1Pr	1 Kit
"Global" Research Assays
"Global" 4-Pop Research	Primate	Low serum, methylcellulose-free	K2-4P-5Pr	1 Kit
"Global" 5-Pop Research	Primate	Low serum, methylcellulose-free	K2-5P-6Pr	1 Kit
"Global" 7-Pop Research	Primate	Low serum, methylcellulose-free	K2-7P-8Pr	1 Kit
"Global" Research Assays
"Global" 4-Pop Research	Primate	Serum-free. Methylcellulose-free	K2SF-4P-5Pr	1 Kit
"Global" 5-Pop Research	Primate	Serum-free. Methylcellulose-free	K2SF-5P-6Pr	1 Kit
"Global" 7-Pop Research	Primate	Serum-free. Methylcellulose-free	K2SF-7P-8Pr	1 Kit

HALO™

The Easy and Rapid In Vitro Research Bioluminescence Assay for Measuring Cell Viability and Proliferation of Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 2	Rat	Low serum. Methylcellulose-free	K2-HPP2-1R	1 Kit
SC-GEMM 1	Rat	Low serum. Methylcellulose-free	K2-GEMM1-1R	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Rat	Low serum. Methylcellulose-free	K2-BFU1-1R	1 Kit
P-BFU 2	Rat	Low serum. Methylcellulose-free	K2-BFU2-1R	1 Kit
P-GM 1	Rat	Low serum. Methylcellulose-free	K2-GM1-1R	1 Kit
P-GM 3	Rat	Low serum. Methylcellulose-free	K2-GM3-1R	1 Kit
P-Mk 1	Rat	Low serum. Methylcellulose-free	K2-Mk1-1R	1 Kit
Lymphopoietic Progenitor Cells
P-Tcell 1	Rat	Low serum. Methylcellulose-free	K2-T1-1R	1 Kit
P-Bcell 1	Rat	Low serum. Methylcellulose-free	K2-B1-1R	1 Kit
No Growth Factors Added
Any	Rat	Low serum. Methylcellulose-free	K2-0-1R	1 Kit
"Global" Research Assays
636"Global" 4-Pop Research	Rat	Low serum, methylcellulose-free	K2-4P-5R	1 Kit
644"Global" 5-Pop Research	Rat	Low serum, methylcellulose-free	K2-5P-6R	1 Kit
652"Global" 7-Pop Research	Rat	Low serum, methylcellulose-free	K2-7P-8R	1 Kit

HALO™

The Easy and Rapid In Vitro Research Bioluminescence Assay for Measuring Cell Viability and ProlifeDogion of Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 2	Dog	Low serum. Methylcellulose-free	K2-HPP2-1C	1 Kit
SC-GEMM 1	Dog	Low serum. Methylcellulose-free	K2-GEMM1-1C	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Dog	Low serum. Methylcellulose-free	K2-BFU1-1C	1 Kit
P-BFU 2	Dog	Low serum. Methylcellulose-free	K2-BFU2-1C	1 Kit
P-GM 1	Dog	Low serum. Methylcellulose-free	K2-GM1-1C	1 Kit
P-GM 3	Dog	Low serum. Methylcellulose-free	K2-GM3-1C	1 Kit
P-Mk 1	Dog	Low serum. Methylcellulose-free	K2-Mk1-1C	1 Kit
No Growth Factors Added
Any	Dog	Low serum. Methylcellulose-free	K2-0-1C	1 Kit
"Global" Research Assays
"Global" 4-Pop Research	Dog	Low serum, methylcellulose-free	K2-4P-5C	1 Kit
"Global" 5-Pop Research	Dog	Low serum, methylcellulose-free	K2-5P-6C	1 Kit
"Global" 7-Pop Research	Dog	Low serum, methylcellulose-free	K2-7P-8C	1 Kit

HALO™

The Easy and Rapid In Vitro Research Bioluminescence Assay for Measuring Cell Viability and ProlifeHorseion of Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 2	Horse	Low serum. Methylcellulose-free	K2-HPP2-1E	1 Kit
SC-GEMM 1	Horse	Low serum. Methylcellulose-free	K2-GEMM1-1E	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Horse	Low serum. Methylcellulose-free	K2-BFU1-1E	1 Kit
P-BFU 2	Horse	Low serum. Methylcellulose-free	K2-BFU2-1E	1 Kit
P-GM 1	Horse	Low serum. Methylcellulose-free	K2-GM1-1E	1 Kit
P-GM 3	Horse	Low serum. Methylcellulose-free	K2-GM3-1E	1 Kit
P-Mk 1	Horse	Low serum. Methylcellulose-free	K2-Mk1-1E	1 Kit
No Growth Factors Added
Any	Horse	Low serum. Methylcellulose-free	K2-0-1E	1 Kit

HALO™

The Easy and Rapid In Vitro Research Bioluminescence Assay for Measuring Cell Viability and ProlifePigion of Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 2	Pig	Low serum. Methylcellulose-free	K2-HPP2-1Po	1 Kit
SC-GEMM 1	Pig	Low serum. Methylcellulose-free	K2-GEMM1-1Po	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Pig	Low serum. Methylcellulose-free	K2-BFU1-1Po	1 Kit
P-BFU 2	Pig	Low serum. Methylcellulose-free	K2-BFU2-1Po	1 Kit
P-GM 1	Pig	Low serum. Methylcellulose-free	K2-GM1-1Po	1 Kit
P-GM 3	Pig	Low serum. Methylcellulose-free	K2-GM3-1Po	1 Kit
P-Mk 1	Pig	Low serum. Methylcellulose-free	K2-Mk1-1Po	1 Kit
No Growth Factors Added
Any	Pig	Low serum. Methylcellulose-free	K2-0-1Po	1 Kit

HALO™

The Easy and Rapid In Vitro Research Bioluminescence Assay for Measuring Cell Viability and ProlifeSheepion of Lympho-Hematopoietic Cells

Cell Population	Species	Formulation	Catalog Number	Quantity
Stem Cells
SC-HPP 2	Sheep	Low serum. Methylcellulose-free	K2-HPP2-1O	1 Kit
SC-GEMM 1	Sheep	Low serum. Methylcellulose-free	K2-GEMM1-1O	1 Kit
Hematopoietic Progenitor Cells
P-BFU 1	Sheep	Low serum. Methylcellulose-free	K2-BFU1-1O	1 Kit
P-BFU 2	Sheep	Low serum. Methylcellulose-free	K2-BFU2-1O	1 Kit
P-GM 1	Sheep	Low serum. Methylcellulose-free	K2-GM1-1O	1 Kit
P-GM 3	Sheep	Low serum. Methylcellulose-free	K2-GM3-1O	1 Kit
P-Mk 1	Sheep	Low serum. Methylcellulose-free	K2-Mk1-1O	1 Kit
No Growth Factors Added
Any	Sheep	Low serum. Methylcellulose-free	K2-0-1O	1 Kit

Please click on the links to take you directly to the videos.

How to Setup HALO®, HemoFLUOR™ or HemoLIGHT™ Hematopoietic Research Assays

How to Calibrate and Standardize Any ATP Bioluminescence Assay - Part 1 (for HALO® Research Assays)

How to Calibrate and Standardize Any ATP Bioluminescence Assay - Part 2 (for HALO® Research Assays)

Please navigate to HemoFLUOR™ or HemoLIGHT™ to see how to perform a fluorescence or absorbance readout. 

 

• Measure lympho-hematopoietic clonal proliferation and cell growth.
• Detect and measure proliferation potential and ability of human and animal stem and progenitor cells, including T-cell progenitor cells.
• Reliably and reproducibly replaces all applications of the colony-forming assay (CFU/CFC).
• Measure and compare proliferation status or potential between tissues and cell populations.
• Experimental transplantation models.
• Effects of growth factors and/or cytokines.
• In vivo to in vitro assays.
• Gene targeting assays, e.g. CRISPR-Cas9.

• HALO® measures the viability, proliferation ability and potential of lympho-hematopoietic cells by using a highly sensitive ATP bioluminescence readout.  
• Correlates directly with any methylcellulose CFC/CFU, fluorescence or absorbance assay.
• Minimal training, simple (no colony counting required), time efficient and cost effective.
• Results are non-subjective, instrument-based and quantitative.
• Fully standardized using external standards and controls included in assay kit.
• Direct comparison of results between samples, and over time without the normalization of data.
• Results in 4 days for animal cells, and 5 days for human cells.
• Incorporates proprietary Suspension Expansion Culture™ (SEC™) Growth Medium to distinguish up to 7 different stem cell, and 9 different progenitor cell populations from 8 species. 
• Also available as 4-, 5- and 7-Population "Global" assays using low serum or serum-free SEC™ growth media.
• HALO® can be multiplexed with other assay readouts such as flow cytometric phenotypic analysis or gene expression assays.

 

 

Please visit the Instructional Videos page

 

Please visit the Instructional Videos page

 

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 instrument and the ATP standard is used to standardize the assay. The values obtained provide the measurement assurance parameters that allow you, the user, to ensure that the results obtained can be trusted.

An example of an ATP standard curve and control results are shown in the diagram below and actual values for the kit being used are given in the Technical Manual. If the values obtained are within the ranges given in the Technical Manual, the user can process the samples knowing that the results will be trustworthy.


 

HALO® is available with a low serum formulation of SEC™ Growth Media for the following species:

• Human
• Non-human primate (Macaca fascicularis (Cynomologus) and Macaca mulatta (Rhesus))
• Horse
• Pig
• Minipig
• Sheep
• Dog
• Rat
• Mouse

HALO® is also available with serum-free SEC™ Growth Media for:

• Human
• Primate
• Mouse

HALO® is available for the following cell populations:
 

Stem Cells

• SC-HPP X: IL-3, IL-6, SCF, Flt3-L (only for human cells and only serum-free)
• SC-HPP 1: IL-3, IL-6, SCF, TPO, Flt3-L
• SC-HPP 2: EPO, GM-CSF, IL-2, IL-3, IL-6, IL-7, SCF, TPO, Flt3-L
• SC-GEMM 1: EPO, GM-CSF, IL-3, IL-6, SCF, TPO, Flt3-L
• SC-GEMM 2: EPO, GM-CSF, IL-3, IL-6, SCF, TPO
• SC-GEM 1: EPO, GM-CSF, IL-3, IL-6, SCF
• SC-GEM 2: EPO, GM-CSF, IL-3, SCF
• SC-GEM 3: EPO, GM-CSF, G-CSF, IL-3, SCF

 

Primitive Stem Cell-Producing, T-Lymphocyte Cell Populations

• SC-HPP XT: IL-3, IL-6, SCF, Flt3-L CD3 CD28 (only for human cells and only serum-free; see also P-Tcells)
• SC-HPP 1T: IL-3, IL-6, SCF, Flt3-L IL-2, CD3 CD28 (only for human cells and only serum-free; see also P-Tcells)

 

Progenitor Cells (P)

• P-BFU 1: EPO, IL-3, SCF
• P-BFU 2: EPO alone
• P-GM 1: GM-CSF, IL-3, SCF
• P-GM 3: GM-CSF alone
• P-Mk 1: TPO, IL-3, SCF

 

Lymphopoietic Cells

• P-Tcell 1: IL-2 alone
• P-Tcell 2: CD3 C28 (human and mouse cells only)
• P-Tcell 3: IL-2, CD3 CD28 (human and mouse cells only)
• P-Bcell 1: Il-7 only (primate, rat, mouse and canine)
• P-Bcell 2: Cocktail includes cytokines only (human cells only)
• P-Bcell 3: Cocktail includes IgD-Dex  cytokines (human and mouse)
• For studying mature immune cells, please refer to ImmunoGlo™, ImmunoGlo™ TCP and ImmunoGlo™ BCP

 

No growth factors: Add growth factors/cytokines of choice.

 

HALO® "Global" Population Assays

 

4-Population "Global" Assays include reagents to measure:

• SC-GEMM 1
• P-BFU 1
• P-GM 1
• P-Mk 1
• Background controls

5-Population "Global" Assays include reagents to measure:

• SC-HPP 2
• SC-GEMM 1
• P-BFU 1
• P-GM 1
• P-Mk 1
• Background control

7-Population "Global" Assays include reagents to measure:

• SC-HPP 2
• SC-GEMM 1
• P-BFU 1
• P-GM 1
• P-Mk 1
• P-Tcell 3
• P-Bcell 3
• Background control

 

Characteristics	HALO:	HemoGLO™	HemoFLUOR™	MultiCellGro™	HemoLIGHT™
Uses:	For all applications where a standardized and validated assay is required. Basic and stem cell research. Veterinary research. Cellular therapy. Hematotoxicity testing and screening	For all hematopoietic viability and cell proliferation applications. Basic and stem cell research. Veterinary research. Limited cellular therapy.	For all hematopoietic viability and cell proliferation applications. Basic and stem cell research. Veterinary research. Limited cellular therapy.	For primitive lympho-hematopoietic stem cell applications. Basic and stem cell research. Veterinary research. Limited cellular therapy.	For all hematopoietic viability and cell proliferation applications. Basic and stem cell research. Veterinary research. Limited cellular therapy.
Assay availability:	1 kit	1 kit	1 kit	1 kit	1 kit
Number of samples:	Depends on number of replicates required(1)	Depends on number of replicates required(1)	Depends on number of replicates required(1)	Depends on number of replicates required(1)	Depends on number of replicates required(1)
Type of assay(2):	Suspension Expansion Culture™ (SEC™)	Suspension Expansion Culture™ (SEC™)	Suspension Expansion Culture™ (SEC™)	Suspension Expansion Culture™ (SEC™)	Suspension Expansion Culture™ (SEC™)
Assay format:	96-well plate. 384-well plate upon request(3)	96-well plate.	96-well plate.	96-well plate	96-well plate
Throughput:	High throughput	High throughput capability	High throughput capability	High throughput capability	High throughput capability
Cell functionality detected:	Cell viability/ cell proliferation/ cytotoxicity	Cell viability/ cell proliferation	Cell viability/ cell proliferation	Cell viability/ cell proliferation	Cell viability/ cell proliferation
Biochemical principle:	Intracellular ATP, luciferin/luciferase	Intracellular ATP, luciferin/luciferase	Resazurin to resorufin (fluorescent)	Resazurin to resorufin (fluorescent)	Tetrazolium WST-1 reduction to yellow formazan
Wavelength (filters):	None. Glow luminescence	None. Glow luminescence	Excitation:560nm. Emission: 590nm	Excitation:560nm. Emission: 590nm	Absorbance at 440nm
Signal development time:	10 minutes	10 minutes	1-3 hours	1-3 hours	1-4 hours
Readout:	Luminescence plate reader(4)	Luminescence plate reader(4)	Fluorescence plate reader (4)	Fluroescence plate reader(4)	Absorbance plate reader(4)
Readout endpoint:	Relative  luminescence units (RLU) with  conversion to ATP concentrations	Relative  luminescence units (RLU) with  optional(5) conversion to ATP concentrations	Relative fluorescence units (RFU)	Relative fluorescence units (RFU)	Relative absorbance units (RAU)
Subjectivity:	Non-subjective	Non-subjective	Non-subjective	Non-subjective	Non-subjective
Cell types detected:	Stem and progenitor cells	Stem and progenitor cells	Stem and progenitor cells	Primitive lympho- hematopoietic stem cells	Stem and progenitor cells
Species:	Human. Primate. Horse. Pig. Sheep. Dog. rat. Mouse	Human. Primate. Horse. Pig. Sheep. Dog. rat. Mouse	Human. Primate. Horse. Pig. Sheep. Dog. rat. Mouse	Human. Primate. Horse. Pig. Sheep. Dog. rat. Mouse	Human. Primate. Horse. Pig. Sheep. Dog. rat. Mouse
MethoCult™ equivalent assays available:	No	Yes	Yes	No	Yes
Total culture volume required (6):	Depends on number of replicates required	Depends on number of replicates required	Depends on number of replicates required	Depends on number of replicates required	Depends on number of replicates required
Suggested number of replicates/culture:	6-8 or as many as required	4-8 or as many as required	4-8 or as many as required	4-8 or as many as required	4-8 or as many as required
Volume/culture:	0.1mL	0.1mL	0.1mL	0.1mL	0.1mL
Cell culture time(7):	3-7 days	4-7 days	4-7 days	4-7 days	4-7 days
Instrument calibration:	Yes(8)	Optional(8)	No	No	No
Standardized assay:	Yes(9)	Optional(9)	No	No	No
Proficiency testing incorporated into assay:	Yes(10)	Optional(10)	No	No	No
Measurement assurance:	Yes(11)	Optional(11)	No	No	No
Validation:	Yes(12)	Optional(12)	No	No	No

(1) = The number of samples tested depends on the number of replicates/sample performed. For example, if 4 replicates/sample are performed, then 24 samples can be analyzed on a single 96-well plate. For 6 replicates, 16 samples. For 8 replicates, 12 replicates. These samples, however, do not have to performed on the same day.
(2) = Suspension Expansion Culture™ (SEC™) Technology takes the place of methylcellulose, allowing easier and faster dispensing.
(3) = Some HALO® assays, including STEMpredict™ and all HALO®-Tox HT assays for hematotoxicity testing are available with 384-well plates upon request.
(4) = Multimode plate readers are also av available that have the capability of luminescence, fluorescence and absorbance.
(5) = HemoGLO™ does not include the reagents to calibrate and standardize the assay. However, these reagents are available separately for this assay.
(6) = The total culture volume will depend on the number of replicates required. For example, if 6 replicates are required, a total volume of 0.8mL should be prepared.
(7) = Culture time depends on the state of fractionation or purification of the source cells, cell population tested and species. In some cases it also depends on the assay itself. For example, STEMpredict™ is a 3-day assay.
(8) = Instrument calibration is performed using the ATP controls included with the assay kit or obtained separately for HemoGLO™.
(9) = Standardization implies performing a standard curve. ATP standards are included in all HALO® assay kits and can be obtained separately for HemoGLO™.. 
10 = The inclusion of standards and controls allows the results to be compared to measurement assurance parameters (provided) to ensure the assay is working correctly. This allows proficiency testing to be determined directly prior to measuring samples. 
11 = Measurement assurance parameters are included in all HALO® assays and when standards and controls are purchased separately for HemoGLO™.
12 = A validated assay allows the user to trust the results obtained. This can only be achieved if standards and controls are included. HALO® and HemoGLO™ have been validated. The validation parameters are included for all HALO® assays and included when the optional standardization kit is purchased for HemoGLO™. Standards and controls allow for in-house validation.

When You Use HALO®, No Colony Counting is Necessary

Since HALO® does not use methylcellulose, no colonies are produced. Instead, HALO® incorporates Suspension Expansion Culture™ (SEC™) Technology; cells are cultured in suspension. This means it is easier, faster and more accurate to setup HALO® cultures because normal pipettes are used. The question always asked is: If no colonies are produced, how do I know whether the cells have differentiated and how do I detect cell differentiation? The answer lies in flow cytometry. The type of cells produced is dependent on the growth factor cocktail used, just as it is in a normal CFU assay. The diagram above show this. If a SC-GEMM culture is setup in HALO®, you would expect to see the presence of stem cells, erythroid cells, granulocytes and macrophages and megakaryocytes, because that is the definition of the SC-GEMM stem cell population. If the cells are removed from a SC-GEMM culture and analyzed by flow cytometry, these are precisely the cell types that are present in the culture as seen by the blue columns. Similarly, if SC-BFU cultures are setup, you would expect to see a prevalence of CD235a+ (glycophorin-A) cells and that is exactly what you find, together with some granulocytes and macrophages as well as megakaryocytes. In other words, the cells you detect in HALO® cultures by flow cytometry are the cells you would expect for the growth factor cocktail used. Therefore, waiting for differentiation to occur and enumerating different types of colonies is unnecessary when using HALO®.

Measuring Proliferation Potential

Proliferation potential is measured by performing a cell dose response and calculating the slope of the linear regression. The steeper the slope, the greater the proliferation potential of the cell population. The greater the proliferation potential, the more primitive the cell population. 

Example of a 7-Population Comparison Between Human Tissues

This graph shows the response of 7 lympho-hematopoietic cell populations derived from mobilized peripheral blood CD34 cells, bone marrow and umbilical cord blood mononuclear cells. Please note that whereas mobilized peripheral blood CD34 cells are shown at 500 CD34 /well, bone marrow and umbilical cord blood are shown at 5,000 mononuclear cells/well.

Comparison of 7-Population Response between Species

The graph shows the response of 7 lympho-hematopoietic populations, SC-HPP 2, SC-GEMM 1, P-BFU 1, P-GM 1, P-Mk 1, P-Tcell, P-Bcell and background, from sheep, pig, dog, rat and mouse bone marrow. 

Production of T-Lymphocytes from Primitive Lympho-Hematopoietic Stem Cells

Human bone marrow High Proliferative Potential-Stem and Progenitor Cells (SC-HPP) were stimulated with various factors and lymphocyte co-factors to induce the cells out of quiecence and into the T-cell lineage. Such studies can be combined with low cytometric phenotypic analysis to determine different T-cell populations. 

For Research Use Only. Not for clinical diagnostic use.

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

• HALO® Master Mix. Please choose between low serum or serum-free formulations.
• ATP standard
• ATP high and low controls (not included with research kits)
• ATP Enumeration Reagent
• Sterile, 96-well plates
• Non-sterile, 96-well plates
• Sterile, adhesive foil covers

Please note that the same protocol is used for HALO® "Global" 4-, 5- and 7-Population Assay Kits

 

Please visit the Instructional Videos page
Please note that Certificate of Analysis (CoAs) will be provided with the assay kit 
Download the HALO® Technical Manual
Download the ATP Optimization Kit Protocol for First-Time Users
Download Luminometer Setup and RLU to ATP Conversion
Download the SEC™ Media SDS sheet
Download the ATP ER SDS sheet
Download the ATP Standard and Controls SDS sheet