X-a-gal

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.

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.