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Sigma-Aldrich

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.

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.
Research Category
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.

Quality

• Each lot is tested by measuring ALDH activity in SK-BR-3 cells by flow cytometry.
• 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.

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

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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Chemosensitization and inhibition of pancreatic cancer stem cell proliferation by overexpression of microRNA-205.
Chaudhary, AK; Mondal, G; Kumar, V; Kattel, K; Mahato, RI
Cancer letters null
Hybrid clone cells derived from human breast epithelial cells and human breast cancer cells exhibit properties of cancer stem/initiating cells.
Gauck, D; Keil, S; Niggemann, B; Zanker, KS; Dittmar, T
BMC Cancer null
Loren E Stallcop et al.
Lab on a chip, 18(3), 451-462 (2018-01-11)
Tape-based razor-printing is a flexible and affordable ultra-rapid prototyping approach for microscale device fabrication. However, integration of this prototyping approach into cell-based assay development has been limited to proof of principle demonstrations. This is in large part due to lack
Natalia Pajuelo-Lozano et al.
Theranostics, 10(21), 9601-9618 (2020-08-31)
Rationale: Gastric cancer (GC) is a solid tumor that contains subpopulations of cancer stem cells (CSCs), which are considered drivers of tumor initiation and metastasis; responsible for therapeutic resistance; and promoters of tumor relapse. The balance between symmetric and asymmetric
Lars Zimmermann et al.
Oncotarget, 8(5), 8173-8188 (2016-12-29)
4-hydroxy-2-nonenal (HNE), a toxic lipid peroxidation product, is associated with oxidative damage in cells and involved in various diseases including the initiation and progression of cancer. Cancer cells have a high, adaptable metabolism with a shift from oxidative phosphorylation to

Articles

AldeRed™ 588-A detects ALDH activity in cancer stem cells, aiding in cancer research.

AldeRed™ 588-A detects ALDH activity in cancer stem cells, aiding in cancer research.

AldeRed™ 588-A detects ALDH activity in cancer stem cells, aiding in cancer research.

AldeRed™ 588-A detects ALDH activity in cancer stem cells, aiding in cancer research.

See All

Protocols

Step-by-step tumorpshere formation protocol that can used to generate 3D cancer stem cell spheroids.

Step-by-step tumorpshere formation protocol that can used to generate 3D cancer stem cell spheroids.

Step-by-step tumorpshere formation protocol that can used to generate 3D cancer stem cell spheroids.

Step-by-step tumorpshere formation protocol that can used to generate 3D cancer stem cell spheroids.

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