PharmaOnco™ ALDH Activity Assay Kit, Microplate

PharmaOnco™ ALDH Activity Assay Kit (Flow Cytometry) is fast and reliable method to quantify the aldehyde dehydrogenase (ALDH) activity. ALDH is a superfamily of phase-I oxidases, which are mainly responsible for the maintenance of cellular homeostasis. It has been reported that ALDH is expressed in stem/progenitor cells, including cancer-initiating cells (CIC) of various organs. This kit provide a quantitative method to detect ALDH enzyme activity in a variety of samples. In this assay, acetaldehyde is oxidized by ALDH generating NADH which then reduces a colorless probe to a colored product with strong absorbance at 450 nm.

Microplate

ALDH assay protocol summary:
1. Add samples and standards to wells;
2. add reaction mix and incubate for 20-60 min;
3. analyze with a microplate reader at 5 min and 20-60 min.

Aldehyde dehydrogenase (ALDH)

Colorimetric

Enzyme activity

Quantitative

Cell culture supernatant, Cell Lysate, Plasma, Serum, Tissue Culture Media

This assay has high sensitivity and excellent specificity for detection of ALDH.

Acetaldehyde, 0.5ml
ALDH Assay Buffer, 1 x 25ml
ALDH Positive Control, 1 vial
ALDH Substrate Mix, 1 vial
NADH Standard, 1 vial

96-well clear plate with flat bottoms
Glycerol
Multi-well spectrophotometer (ELISA reader)
Pipettes

0.5 mU/well

30 min

1. Liquid samples can be measured directly. Tissue (50 mg) or cells (1 x 106) should be rapidly homogenized with ~ 200 µl ice cold ALDH Assay Buffer for 10 minutes on ice.
2. Spun down at 12000 rpm for 5 min to remove nuclei and insoluble material.
3. Add 1- 50 ul of the collected supernatant into a 96 well plate and adjust the final volume to 50 µl with ALDH Assay Buffer.
5. Prepare a total 50 µl Reaction Mix containing: ALDH Assay Buffer 43 µl, ALDH Substrate Mix 2 µl, Acetaldehyde 5 µl.
6. Add 50 µl of the Reaction Mix to each well.
7. Incubate at room temperature for 5 min and measure the OD of samples and sample backgrounds at 450 nm.

Mammals, Other species

Store at -20°C

Stable as supplied until expiry date on label.

Restrictions for research use only. Intended for use by laboratory professionals.