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Products - HemoFLUOR

  •  HemoFLUOR™-96

  •  HemoFLUOR™-96 PCAEQ

  •  HemoFLUOR™-96 PMT

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HemoFLUOR™-96

Fluorescence Viability and Proliferation Assays for Lympho-Hematopoietic Stem and Progenitor Cells

 

Buy HemoFLUOR™-96 Products

Available as assay kits or
contract research services

 

 

HemoFLUOR™-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 HemoFLUOR™-96
  • No colony counting needed.
  • Non-subjective, instrument-based and quantitative.
  • Uses a plate reader or multiparameter plate reader capable of measure fluorescence (380-400nm excitation/505nm emission).
  • Multiplexes with flow cytometry allowing cells to be detected using other fluorescent labels/markers. 
  • Multiplexes with bioluminescence assays.
  • Use in place of colorimetric/absorbance assays for greater sensitivity.
  • Results in about 7 days depending on cell population and species.
  • Complete assay kit includes cell population-specific HemoFLUOR™ Master Mix and cell processing reagent.
  • Easy to learn, fast to use and greater flexibility.
  • HemoFLUOR™-96 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 HemoFLUOR™-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 HemoFLUOR™-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 HemoFLUOR™-96

5-Population Assay

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

 

HemoFLUOR™-96 Principle
  • Incorporates a customized Promega CellTiter Fluor™ reagent that detects protease activity only in living cells. Fluorescence produced only when the GF-AFC substrate is cleaved by protease activity to produce a fluorescence signal proportional to the number of living cells.
  • After culture, just add 100μl of the reagent, mix and measure signal between 30 minutes and 3 hours in a plate reader. Optimal incubation time: 2 hours.
  • Uses a plate fluorometer or multiparameter plate reader to measure fluorescence with an excitation filter of 380-400nm and an emission filter of 505nm.  

 

Use

For Research Use Only.

 

Major Equipment Required

A fluorescence plate reader or multiparameter plate reader with an excitation filter at 390-400nm and an emission filter at 505nm.

 

Kit Contents
  • HemoFLUOR™-96 Master Mix
  • GF-AFC Reagent
  • Sterile 96-well culture plates
  • Sterile adhesive foil covers to keep unused wells sterile
  • Instruction manual

 

Related Assays

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HemoFLUOR™-96 PCAEQ

Hematopoietic Progenitor Colony-Forming Unit (CFU) Equivalent Fluorescence Assays for Stem Cell Processing Laboratories

 

Buy HemoFLUOR™-96 PCAEQ Products

Available as assay kits or
contract research services

 

HemoFLUOR™-96 PCAEQ 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 HemoFLUOR™-96 PCAEQ in the Stem Cell Processing Laboratory
  • No colony counting required.
  • Transforms the subjective CFU assay into an instrument-based, quantitative assay.
  • 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.
  • Results in 7 days.
  • Faster and easier to use than any traditional CFU assay, even using automated colony counting.
  • Complete assay kit; includes everything needed to culture and obtain results.
  • Easy to learn, fast to use and cheaper than the methylcellulose CFU assays, especially MethoCult® Express.
  • HemoFLUOR™-96 PCAEQ incorporates the same proven Suspension Expansion Culture™ (SEC™) Technology used in HALO® assays.

 

Tissues
  • HemoFLUOR™-96 PCAEQ can be used for the following tissues:
  • Mobilized peripheral blood
  • Umbilical cord blood
  • Bone marrow
  • Purified cell populations for any of the above tissues

 

HemoFLUOR™-96 PCAEQ 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 HemoFLUOR™-96 PCAEQ 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.

 

CAMEO™-4 and HemoFLUOR™-96 PCAEQ Assays that are Equivalent to MethoCult® Reagents
CAMEO™-4 HemoFLUOR™-96 PCAEQ MethoCult® Growth Factor/Cytokine Cocktail
KC-GEM2-50H K5-PCA1-1,2,4* H4434 EPO, GM-CSF, IL-3, SCF
KC-GEM3-50H K5-PCA2-1,2,4* H4034 EPO, GM-CSF, G-CSF, IL-3, SCF
K2-GM1-50H K5-PCA3-1,2,4* H4534 GM-CSF, IL-3, SCF
KC-GM2-50H K5-PCA4-1,2,4* H4035 GM-CSF, G-CSF, IL-3, SCF

*denotes that assay kits are available with 1, 2 or 4, 96-well plates
 
HemoFLUOR™-96 PCAEQ Principle
  • Incorporates a customized Promega CellTiter Fluor™ reagent that detects protease activity only in living cells. Fluorescence produced only when the GF-AFC substrate is cleaved by protease activity to produce a fluorescence signal proportional to the number of living cells.
  • After culture, just add 100μl of the reagent, mix and measure signal between 30 minutes and 3 hours in a plate reader. Optimal incubation time, approx. 2 hours.
  • Uses a plate fluorometer or multiparameter plate reader to measure fluorescence with an excitation filter of 380-400nm and an emission filter of 505nm.

 

Use

For Research Use Only. Not for clinical diagnostic use.

 

Major Equipment Required

A fluorescence plate reader or multiparameter plate reader with an excitation filter at 390-400nm and an emission filter at 505nm.

 

Kit Contents
  • HemoFLUOR™-96 PCAEQ Master Mix
  • GF-AFC reagent
  • Sterile, 96-well plate(s)
  • Sterile, adhesive foil covers so that unused wells remain sterile
  • Instruction Manual

 

Related Assays

 

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HemoFLUOR™-96 PMT

Fluorescence Assays for Hematopoietic Patient Monitoring After Transplantation

 

Buy HemoFLUOR™-96 PMT Products

Available as assay kits or
contract research services

 

 

HemoFLUOR™-96 PMT Applications

Neutrophil, platelet and red blood cell engraftment assays by detecting 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 to using HemoFLUOR™-96 PMT for Patient Monitoring
  • 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.
  • Transforms the subjective CFU assay into a on-subjective, instrument-based, quantitative assay.
  • Easier to use than any traditional CFU assay, even using automated colony counting.
  • Complete assay kit; includes everything needed to culture and obtain results.
  • Easy to learn, fast to use and cheaper than the methylcellulose CFU assays.
  • HemoFLUOR™-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

 

HemoFLUOR™-96 PMT Principle
  • Incorporates a customized Promega CellTiter Fluor™ reagent that detects protease activity only in living cells. Fluorescence produced only when the GF-AFC substrate is cleaved by protease activity to produce a fluorescence signal proportional to the number of living cells.
  • After culture, just add 100μl of the reagent, mix and measure signal between 30 minutes and 3 hours in a plate reader. Optimal incubation time, approx. 2 hours.
  • Uses a plate fluorometer or multiparameter plate reader to measure fluorescence with an excitation filter of 380-400nm and an emission filter of 505nm.

 

Uses

For research use only. Not for clinical diagnostic use.

 

Major Equipment Required

A fluorescence plate reader or multiparameter plate reader with an excitation filter at 390-400nm and an emission filter at 505nm.

 

Kit Contents
  • HemoFLUOR™-96 PMT Master Mix
  • GF-AFC Reagent
  • Sterile, 96-well plate(s)
  • Sterile, adhesive cover foils to maintain sterility of unused wells
  • Instruction manual

Related Assays

Hemogenix Inc.