SCR150
AldeRed® ALDH Detection Assay
Synonym(s):
ALDH Assay, Aldehyde Dehydrogenase Assay, Cancer Stem Cell Assay
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About This Item
UNSPSC Code:
12352207
eCl@ss:
32161000
NACRES:
NA.84
technique(s)
cell culture | stem cell: suitable
flow cytometry: suitable
Quality Level
fluorescence
λex 488 nm
λex 532 nm
λem 615 nm
detection method
fluorometric
shipped in
wet ice
General description
The AldeRed ALDH Detection Kit provides cancer and stem cell scientists with new capabilities for live cell isolation and characterization. The AldeRed reagent is a red-shifted fluorescent substrate for aldehyde dehydrogenase (ALDH), allowing cells to be identified and isolated by flow cytometry with concurrent use of green fluorescent cell lines, antibodies, transgenic animals and reporter assays.
Learn more about the AldeRed in Nature Communications! (Learn Now!)
High ALDH activity serves as a universal marker of stem cells, both normal and malignant. Cells can be identified and isolated based upon the enzymatic activity of ALDH, a detoxifying enzyme responsible for oxidation of hazardous aldehyde byproducts. The marker ALDH has been used to isolate cancer stem cells from various human malignancies including bladder, breast, cervical, colon, head and neck, liver, lung, pancreas, prostate and ovary. The AldeRed assay employs a fluorescent and non-toxic ALDH substrate (AldeRed 588-A) 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 assessement.
Learn more about the AldeRed in Nature Communications! (Learn Now!)
High ALDH activity serves as a universal marker of stem cells, both normal and malignant. Cells can be identified and isolated based upon the enzymatic activity of ALDH, a detoxifying enzyme responsible for oxidation of hazardous aldehyde byproducts. The marker ALDH has been used to isolate cancer stem cells from various human malignancies including bladder, breast, cervical, colon, head and neck, liver, lung, pancreas, prostate and ovary. The AldeRed assay employs a fluorescent and non-toxic ALDH substrate (AldeRed 588-A) 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 assessement.
Application
• For optimal excitation and emission of AldeRed 588-A, use a combination of blue laser (488 nm) or green laser (532 nm) with PE-Texas Red detector (615 nm). Detection in proximal measurement channels (e.g. PE) is also possible, but may affect delta MFI (mean fluorescence intensity) between Test sample and DEAB-treated control sample.
• All live intact cells (even with low ALDH expression) become fluorescent upon exposure to activated AldeRed 588-A. The generation of a DEAB-treated control is necessary for each cell sample to establish baseline fluorescence.
• Handle all reagents using aseptic techniques.
• All live intact cells (even with low ALDH expression) become fluorescent upon exposure to activated AldeRed 588-A. The generation of a DEAB-treated control is necessary for each cell sample to establish baseline fluorescence.
• Handle all reagents using aseptic techniques.
Research Category
Stem Cell Research
Apoptosis & Cancer
Stem Cell Research
Apoptosis & Cancer
Components
1. AldeRed Reagent: (Part No. CS216596). One (1) vial containing 50 µg of AldeRed 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. AldeRed Assay Buffer: (Part No. CS216592). Four (4) bottles containing 25 mL of AldeRed Assay Buffer each.
6. Verapamil: (Part No. CS220012). Four (4) vials containing 615 µg Verapamil powder each.
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. AldeRed Assay Buffer: (Part No. CS216592). Four (4) bottles containing 25 mL of AldeRed 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.
• AldeRed® Reagent is 95% pure (PMR)
• AldeRed® Reagent is 95% pure (PMR)
Storage and Stability
• Store kit at 2-8°C.
• Reconstituted and activated AldeRed Reagent may be stored in aliquots at -20°C protected from light for up to 1 year. Repeated freezing and thawing is not recommended.
• AldeRed Assay Buffer supplemented with Verapamil may be stored at 2-8°C in aliquots for up to 3 months.
• Reconstituted and activated AldeRed Reagent may be stored in aliquots at -20°C protected from light for up to 1 year. Repeated freezing and thawing is not recommended.
• AldeRed Assay Buffer supplemented with Verapamil may be stored at 2-8°C in aliquots for up to 3 months.
Legal Information
ALDERED is a registered trademark of Merck KGaA, Darmstadt, Germany
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
Flash Point(F)
55.4 °F
Flash Point(C)
13 °C
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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Find documentation for the products that you have recently purchased in the Document Library.
Targeting cancer stem cell propagation with palbociclib, a CDK4/6 inhibitor: Telomerase drives tumor cell heterogeneity.
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Articles
AldeRed™ 588-A is a red fluorescent live cell probe that detects ALDH activity used to identify cancer stem cells and progenitor cells in culture. Cancer stem cells (CSCs) are subpopulations of cancer cells that can self-renew, generate diverse cells in the tumor mass, and sustain tumorigenesis. Some researchers believe that cancer arises from cancer stem cells that originate as a result of mutational hits on normal stem cells.
Cancer stem cell media, spheroid plates and cancer stem cell markers to culture and characterize CSC populations.
Protocols
Step-by-step tumorpshere formation protocol that can used to generate 3D cancer stem cell spheroids.
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