HemoLIGHT™-96

Colorimetric / Absorbance Assays for Hematopoietic Stem and Progenitor Cells

Buy HemoLIGHT-96 Products

Available as assay kits or
contract research services

HemoLIGHT™-96 Applications

• Determine the number of viable cells in proliferation.
• Compare hematopoietic cell populations from different species.
• Replaces hematopoietic colony-forming cell (CFC) assay applications.
• Experimental transplantation models.
• Effects of growth factors/cytokines.
• In vivo to in vitro assays.
• Gene targeting assays.

Advantages of using HemoLIGHT™-96 for your Research

• Uses a colorimetric/absorbance ELISA plate reader, availble in most laboratories.
• No colony counting required.
• Non-subjective, instrument-based and quantitative.
• Results in about 7 days depending on species.
• Complete assay kit; includes everything needed to culture and measure hematopoietic cell growth.
• Easy to learn, fast to use and cheaper than the methylcellulose CFC assay.
• HemoLIGHT™ incorporates the same proven Suspension Expansion Culture™ (SEC™) Technology developed for HALO® bioluminescence assays.

Species

• Human
• Non-human primate
• Dog
• Rat
• Mouse

Individual Human Cell Populations Detected using HemoLIGHT™-96

(see Lympho-Hematopoietic System to view cell populations and growth factor/cytokine cocktails)
The following proliferation status or potential can be detected for individual populations using HemoLIGHT™-96:
Stem Cells

• HPP-SP 2: EPO, GM-CSF, IL-2, IL-3, Il-6, IL-7, SCF, TPO, Flt3-L
• CFC-GEMM 1: EPO, GM-CSF, IL-3, IL-6, SCF, TPO, Flt3-L
• CFC-GEM 1: EPO, GM-CSF, IL-3, IL-6, SCF
• CFC-GEM 2: EPO, GM-CSF, IL-3, SCF

Progenitor Cells

• BFU-E 1: EPO, IL-3, SCF
• GM-CFC 1: GM-CSF, IL-3, SCF
• Mk-CFC 1: TPO, IL-3, SCF

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

Multiple Human Cell Populations Detected Simultaneously Using HemoLIGHT™-96 5-Population Assay

• HPP-SP 2
• CFC-GEMM 1
• BFU-E 1
• GM-CFC 1
• Mk-CFC 1
• Background control 

The HemoLIGHT™-96 Principle

• Incorporates a customized Promega CellTiter 96® AQueous One reagent that measures the amount of soluble formazan by absorbance at 490nm produced by a MTS tetrazolium reduction, colorimetric reaction.
• After culture, just add 20μl of the MTS reagent, mix and read absorbance at 490nm after 1-4 hours in a 96-well plate reader. Plates can be returned to incubator and re-read at a later time for increased sensitivity.
• No solubilization step as in a MTT reaction. Replaces MTT assays.
• Read results between 1-4 hours after addition of MTS reagent. Plates can be returned to incubator and re-read at a later time for increased sensitivity.

Use

For Research Use Only.

Major Equipment Required

An absorbance plate readr or multiparameter plate reader to measure absorbance at 490nm.

Kit Contents

• HemoLIGHT™-96 Master Mix
• Sterile 96-well culture plates
• Sterile adhesive foil covers to keep unused wells sterile.
• Instruction manual

Related Assays

• HemoFLUOR™-96
• HALO®-96 Research
• HALO®-96 PREP

HemoLIGHT™-96 PCA^EQ

Colony-Forming Unit (CFU) Equivalent Colorimetric / Absorbance Assays

Buy HemoLIGHT-PCA^EQ Products

Available as assay kits or
contract research services

HemoLIGHT™-96 PCA^EQ Applications

• Determine the number of viable cells in proliferation.
• An alternative to, or replacement for the classic colony-forming unit (CFU) assay for stem cell processing laboratories.
• FDA, FACT and AABB alternative assays. (NMDP uses FACT and AABB standards).

Advantages of using HemoLIGHT-96 in the Stem Cell Processing Laboratory

• No colony counting required.
• Transforms the subjective CFU assay into an instrument-based, quantitative assay.
• Uses an absorbance ELISA plate reader available in most laboratories.
• Incorporates the same growth factor cocktails used in MethoCult® reagents for direct assay-to-assay comparisons and correlation.
• Non-subjective, instrument-based and quantitative assay system.
• Results in 7 days depending.
• Faster and easier to use than any traditional CFU assay, even using automated colony counting.
• Cheaper than the methylcellulose CFU assays.
• Complete assay kit; includes everything needed to culture and obtain results.
• Flexibility to obtain results depending on work schedule.
• HemoLIGHT™-96 PCAEQ incorporates the same proven Suspension Expansion Culture™ (SEC™) Technology used in HALO® assays.

Tissues

HemoLIGHT-96 PCA^EQ can be used for the following tissues:

• Mobilized peripheral blood
• Umbilical cord blood
• Bone marrow
• Purified cell populations for any of the above tissues

HemoLIGHT™-96 PCA^EQ is used as an Alternative to or Replacement for CAMEO™-4

 (using the same reagent) and MethoCult® (using the same growth factor cocktails)

The table below shows the HemoLIGHT™-96 PCA^EQ assays that are equivalent to CAMEO™-4 assays and MethoCult® reagents used in the stem cell processing laboratory. The growth factor/cytokine cocktail is the same for all three.

HemoLIGHT™-96 PCA^EQ Principle

• Incorporates a customized Promega CellTiter 96® AQueous One reagent that measures the amount of soluble formazan by absorbance at 490nm produced by a MTS tetrazolium reduction, colorimetric reaction.
• After culture, just add 20μl of the MTS reagent, mix and read absorbance at 490nm after 1-4 hours in a 96-well plate reader.
• No solubilization step as in a MTT reaction.
• Read results between 1-4 hours after addition of MTS reagent. Plates can be returned to incubator and re-read at a later time for increased sensitivity.

Use

Major Equipment Required

Kit Contents

• HemoLIGHT™-96 PCA^EQ Master Mix
• MTS reagent
• Sterile, 96-well plate(s)
• Sterile, adhesive cover foils to maintain sterility of unused wells
• Instruction manual

Related Assays

• HemoFLUOR™-96 PCA^EQ
• HALO®-96 PCA^EQ
• CAMEO4 for Stem Cell Processing Laboratories 

HemoLIGHT™-96 PMT

Colorimetric / Absorbance Assays for Patient Monitoring after Transplantation

Buy HemoLIGHT-96 PMT Products

Available as assay kits or
contract research services

HemoLIGHT™-96 PMT Applications

• Neutrophil, platelet and red blood cell engraftment assays by detection of GM-CFC, Mk-CFC and BFU-E proliferation and growth.
• Determine hematopoietic stem and progenitor cell reconstitution.
• An alternative to, or replacement for the classic colony-forming unit (CFU) assay for hematopoietic lineage engraftment and reconstitution.

Advantages of using HemoLIGHT™-96 PMT in the Stem Cell Processing Laboratory

• Measurement of progenitor cell proliferation, rather than differentiation, provides faster results.
• Measures individual hematopoietic lineages providing a faster prediction of possible engraftment than the “classic” CFU assay.
• No colony counting required.
• Results in 7 days.
• Transforms the subjective CFU assay into an instrument-based, quantitative assay.
• Easier to learn and faster to use than any traditional CFU assay, even using automated colony counting.
• Complete assay kit; includes everything needed to culture and measure hematopoietic cell growth.
• HemoLIGHT™-96 PMT incorporates the same proven Suspension Expansion Culture™ (SEC™) Technology used in HALO® assays.

Tissues

• Peripheral blood
• Bone marrow (biopsy)

Individual Cell Populations Detected

• GM-CFC to predict neutrophil engraftment
• Mk-CFC to predict platelet engraftment
• BFU-E to predict red blood cell engraftment

Multiple Lineages for "Global" Hematopoietic Reconstitution

• HPP-SP, primitive stem cell
• CFC-GEMM, hematopoietic stem cell
• GM-CFC, myelomonocytic lineage
• BFU-E, erythropoietic lineage
• Mk-CFC, megakaryopoietic lineage

HemoLIGHT™-96 PMT Principle

• Incorporates a customized Promega CellTiter 96® AQueous One reagent that measures the amount of soluble formazan by absorbance at 490nm produced by a MTS tetrazolium reduction, colorimetric reaction.
• After culture, just add 20μl of the MTS reagent and read absorbance at 490nm after 1-4 hours in a 96-well plate reader.
• Plates can be returned to incubator and re-read at a later time for increased sensitivity.
• No solubilization step as in a MTT reaction.

Use

For Research Use Only. Not for clinical diagnostic use.

Major Equipment Required

An absorbance plate readr or multiparameter plate reader to measure absorbance at 490nm.

Kit Contents

• HemoLIGHT™-96 PMT Master Mix
• MTS reagent
• Sterile, 96-well plate(s)
• Sterile adhesive cover foils for maintaining unused well sterile
• Instruction manual

Related Assays

• HemoFLUOR™-96 PMT
• HALO®-96 PMT
• CAMEO™-4 for Stem Cell Processing Laboratories