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technique(s)
flow cytometry: suitable
suitability
suitable for fluorescence
detection method
fluorometric
General description
The AldeGreen ALDH Detection Assay enables flow cytometry-based detection of intracellular Aldehyde Dehydrogenase (ALDH) enzyme activity. High ALDH activity serves as a universal marker of stem cells, both normal and malignant. The assay employs a fluorescent and non-toxic ALDH substrate (AldeGreen) that diffuses freely into intact and viable cells, but remains trapped inside the cells once converted by ALDH into the corresponding acid. The amount of fluorescence produced is proportional to the ALDH activity in the cells and is measured by flow cytometry, allowing fluorescence-activated cell sorting (FACS). This kit supplies the ALDH inhibitor diethylaminobenzaldehyde (DEAB), which is used in negative control testing necessary for background fluorescence assessment.
Application
- AldeGreen-positive cells may be imaged using a fluorescence microscope or quantified using a flow cytometer. A 488 nm argon ion laser may be used for excitation and an optical filter set to 515-545 nm emission for detection. <br />
- All live intact cells (even with low ALDH expression) become fluorescent upon exposure to activated AldeGreen. The generation of a DEAB-treated control is necessary for each cell sample to establish baseline fluorescence.<br />
- Handle all reagents using aseptic techniques.
Components
1. AldeGreen Reagent: (Part No. CS226325). One (1) vial containing 50 µg of AldeGreen reagent powder. 2. DEAB Reagent: (Part No. CS216595). One (1) vial containing 1 mL of DEAB (Diethylaminobenzaldehyde) reagent. 3. Hydrochloric Acid (2N): (Part No. CS216594). One (1) vial containing 1.5 mL of 2 Normal hydrochloric acid. 4. Dimethylsulphoxide: (Part No. CS216593). One (1) vial containing 1.5 mL of Dimethylsulphoxide (DMSO). 5. AldeGreen Assay Buffer: (Part No. CS226324). Four (4) bottles containing 25 mL of AldeGreen Assay Buffer each. 6. Verapamil: (Part No. CS220012). Four (4) vials containing 615 µg Verapamil powder each.
Quality
- Each lot is tested by measuring ALDH activity in SK-BR-3 cells by flow cytometry.
- AldeGreen Reagent is 95% pure (PMR)
Storage and Stability
- Store kit at 2-8°C. • Reconstituted and activated AldeGreen Reagent may be stored in aliquots at -20°C protected from light for up to 1 year. Repeated freezing and thawing is not recommended.• AldeGreen Assay Buffer supplemented with Verapamil may be stored at 2-8°C in aliquots for up to 3 months.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Acute Tox. 3 Oral - Aquatic Chronic 2 - Eye Irrit. 2 - Flam. Liq. 2 - Met. Corr. 1
Storage Class Code
3 - Flammable liquids
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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