ALDEFLUOR™ Assay Buffer (Catalog #01701)
- ALDEFLUOR™ Assay Buffer, 25 mL
ALDEFLUOR™ Assay Buffer (Catalog #01702)
- ALDEFLUOR™ Assay Buffer, 55 mL
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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