For the identification, evaluation, and isolation of stem and progenitor cells expressing high levels of ALDH
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High ALDH expression has been reported for normal and cancer precursor cells of various lineages. The ALDEFLUOR™ assay is a widely published non-immunological method for the detection of ALDH-bright (ALDHbr) cells and can be used to detect cancer cells in hematopoietic, mammary, endothelial, mesenchymal, neural, and other tissues.
ALDEFLUOR™ DEAB Reagent and ALDEFLUOR™ Assay Buffer, included with the kit, support the optimal performance of the assay and are also available for purchase individually.
The kit is compatible with standard flow cytometers for performing downstream analysis of ALDHbr cells and with standard cell sorters for further purification and characterization.
View our additional resources to learn more about the ALDEFLUOR™ reagent system.
Figure 1. Identification of ALDHbr Cells from Mouse Embryonic Brain Samples
E14 SVZ cells stained with ALDEFLUOR™. FACS profiles of DEAB control (A) and ALDH staining (B).
Figure 2. Identification of ALDHbr SSC LO Cells from Human Hematopoietic Samples
Bone marrow low density cells (A-B), peripheral blood mononuclear cells (C, D) and umbilical cord blood cells (E, F) stained with ALDEFLUOR™. FACS profiles of DEAB control (A, C, E) and ALDH staining (B, D, F).
Figure 3. Identification of ALDHbr Cells from Human Breast Cancer Cell Lines
SKBR3 breast cancer cells stained with ALDEFLUOR™ for 45 minutes. FACS profiles of DEAB control (A) and ALDH staining (B).
Figure 4. Identification of ALDHbr Cells from Human Mammary Epithelial Samples
Primary normal human mammary epithelial samples stained with ALDEFLUOR™. FACS profiles of DEAB control (A) and ALDH staining (B).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (23)
Frequently Asked Questions
The reagents in the kit were frozen when I received it. Will this cause a problem?
Is it acceptable for activation of the ALDEFLUOR™ reagent to exceed 30 minutes?
Can I speed up the activation reaction by incubating at 37°C?
Will the activation reaction proceed at refrigerator (2 - 8°C) temperatures?
How should I store the ALDEFLUOR™ reagent after it is activated?
Why must the ALDEFLUOR™ assay buffer be added?
Is it acceptable for the staining reaction to exceed 30 minutes?
Will the staining reaction proceed at refrigerator (2 - 8°C) temperatures?
Can I add any other efflux inhibitors to the ALDEFLUOR™ assay buffer?
• 50 - 100 µM verapamil
• 2.5 mM probenecid
• 100 mM 2-deoxy-D-glucose
• 1 mg/mL sodium azide (0.1%) Note: Sodium azide may be toxic to cells. Do not use if cellular function assays are to be performed after the ALDEFLUOR™ assay.
Note: Ice is the universal efflux inhibitor. Keep all ALDEFLUOR™-reacted samples on ice or at 2 - 8°C as much as possible.
Can I stain the cells at a concentration higher than 1 x 106 cells/mL?
What anticoagulants can be used to collect samples?
Do erythrocytes (red blood cells) interfere with the assay?
What solutions can be used to lyse erythrocytes?
• Ammonium chloride (e.g. 0.17 M NH4CI, 10 mM Tris HCI, 0.25 mM EDTA),
• 1X ABC Lysis Buffer (eBioscience, San Diego, CA)
• VitaLyse® (BioE, St Paul, MN).
We do not recommend use of the following or any other solution that contains a fixative, as these will render the cells nonviable:
• CyLyse® (Partec GMBH, Munster, Germany),
• FACS™ Lysing solution (BD Biosciences, San Jose, CA.)
Can fixed cells be used with this assay?
Does the ALDEFLUOR™ assay work on cryopreserved cells?
Will ALDEFLUOR™ buffer prevent efflux in cells from non-hematopoietic tissues or from other species?
Will DEAB inhibit ALDH activity in cells from non-hematopoietic tissues or from other species?
Can I use a greater concentration of the ALDEFLUOR™ substrate to improve the discrimination of the ALDHbr population?
Can I analyze cells by the ALDEFLUOR™ assay and the side population assay simultaneously?
Why are all the cells in the cytogram fluorescent to some degree?
How do I compensate for multiparameter flow analysis when the staining of ALDHbr cells is so bright?
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ALDEFLUOR is registered trademark of ALDAGEN Inc.
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.