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SCR150 Sigma-Aldrich

AldeRed ALDH Detection Assay

  •  eCl@ss 32161000

  •  NACRES NA.84

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Properties

Quality Level   100
application(s)   cell culture | stem cell: suitable
  flow cytometry: suitable
fluorescence   λex 488 nm
  λex 532 nm
  λem 615 nm
detection method   fluorescent
shipped in   wet ice

Description

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.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

Cancer Stem Cell Detection Utilizing a Red-shifted Fluorescent Live Cell Probe for Aldehyde Dehydrogenase (ALDH), AldeRed™ 588-A.

Introduction Methods    Reagent Preparation    Stem Cell Sample Preparation    AldeRed ALDH Assay    Flow Cytometry Analysis Results Materials
Nick Asbrock1, Konstantin Taganov1, Vi Chu1, Martin G. Pomper2, Il Minn2, Haofan Wang2, Steven D. Leach2

1Assay & Platform Development, Bioscience BU, MilliporeSigma, 28820 Single Oak Drive, Temecula, CA, USA 9259
2Johns Hopkins Russel H. Morgan Department of Radiology and Radiological Science, Baltimore, Maryland 21287, USA.
Keywords: Cancer, Enzyme activity, Evaporation, Flow cytometry, Oxidations, Sample preparations

Cancer Stem Cells: New Targets for Cancer Therapy

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Keywords: Cancer, Cell attachment, Cell culture, Cell proliferation, Gastrointestinal, Gene expression, Transcription, transformation

Protocols

Cancer Stem Cell Tumorsphere Formation Protocol

Solid tumors grow in a three-dimensional (3D) spatial conformation, resulting in a heterogeneous exposure to oxygen and nutrients as well as to other physical and chemical stresses. To mimic the 3D s...
Keywords: Cancer, Cell culture

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Hematopoietic Stem and Immune Cell Culture

Hematopoietic stem cells (HSCs) give rise to all blood cell types including myeloid and lymphoid lineages. Myeloid cells include monocytes, macrophages, neutrophils, basophils, eosinophils, erythrocy...
Keywords: Cell biology, Cell culture, Centrifugation, Clinical, Culture media, Flow cytometry, Growth factors, Ligands, Stem cell biology

Peer-Reviewed Papers
15

References

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