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CAS:107021-38-5
Appearance:White Powder
Storage:Store at -20℃，3 years
Purity:95.00%
Product Information:
X-alpha-gal is a chromogenic substrate for yeast galactosidase (MEL1), which is used to directly detect yeast two-hybridization in the GAL4 system on medium. Rapid and easy identification of positive blue clones by blue-white color screening without time-consuming β-galactosidase reporter gene testing.
X-alpha-gal assays the activation of the yeast MEL1 gene, a reporter gene in the GAL4 yeast two-hybrid system that encodes secretory alpha-galactosidase. The enzyme hydrolyzes the colorless X-alpha-gal substrate and eventually produces a blue product. Yeast expressing alpha-galactosidase land blue on a medium containing X-alpha-gal, i.e. positive double hybridization.
x-α-gal A-galactosidase chromogenic substrate Use:
(1) The chromogenic substrate of α-galactosidase, forming a blue precipitate. The yeast two-hybrid interaction was directly detected on the medium, and the β-galactosidase solution and filter-lift assays in the two-hybrid system could be eliminated by using this substrate.
(2) Used to distinguish different strains of coliaceae and bifidobacteria and Lactobacillus wild-type and galactosidase gene defective strains;
(3) Used in histochemistry for the detection of enzyme activity.
Preparation and use of storage solution:
A) Coated on a prefabricated plate: dissolve 24mgX-α-gal in a solution of 6mLDMF with a final concentration of 4mg/mL.
Apply 200ul(15cm) or 100ul (10cm) X-alpha-Gal storage solution to a prefabricated plate;
② Put in the incubator at 37℃until the liquid is absorbed (up to 4 hours due to low DMF volatility);
③ The transformed bacteria or yeast were coated on a plate and cultured at 37℃or 30℃until blue colonies appeared.
B) Directly added to AGAR: dissolve 60mg X-α-gal in 3mLDMF solution with a final concentration of 20mg/ mL.
① The sterilized AGAR medium was cooled to 50-55℃;
② Directly add 2ml-10ml20mg/mlX-α-Gal storage solution to the cooled medium, mix well, and quickly pour onto the plate.
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