Gus Histochemical Stain Kit

Gus Histochemical Stain Kit

GUS Staining Kits

GUS003

GUS is a very comprehensive reporter gene detection system widely used in plant molecular biology and microbiology research. The GUS gene fusion system can qualitatively or quantitatively analyze the GUS enzymatic activity of cells/tissues or protein extracts according to the type of substrate. Histochemical methods (qualitative studies) are used for subcellular localization of GUS fusion protein expression, mostly using X-Gluc as a substrate, because it can be hydrolyzed to generate a colorless glucuronic acid and a visible nitrogen-bromoindigo ( chloro-tromoindigo) dark blue precipitate. Fluorescence analysis (fixation research) is used to study the expression level of GUS fusion proteins, often using MUG as a substrate, because it can be hydrolyzed to generate a colorless glucuronic acid and a fluorescent product 4-MU (4-methyl) base umbelliferone), blue fluorescence (Ex=365 nm, Em=445 nm). Since there is no endogenous GUS activity in most plant cells, GUS is widely used as a reporter gene for transgenic plants, especially in transformation experiments for transient expression of exogenous genes.

Localization (Qualitative) Study of GUS Expression in Transgenic Plants.

This kit uses X-Gluc as the chromogenic substrate for histochemical staining of GUS fusion protein (qualitative research). In addition, the kit contains all the reagents required for staining, and the GUS staining working solution can be prepared by mixing X-Gluc solution and GUS staining buffer in proportion. This kit can prepare 50 mL of staining working solution. It has the advantages of simple, fast, stable operation and low background.

• X-Gluc Substrate, 1 Vial.
• X-Gluc Solvent Substrate Solution, 1 Vial.
• Gus Stain Buffer GUS staining buffer, 50 mL.

Dry ice.

Store at 4 ℃ for 12 months.

GUS staining procedure
(1) Materials: Cut rhizomes, leaves, petals and other tissues into small pieces and put them in a 1.5ml centrifuge tube.
Note: The method of preparation of plant material for staining varies depending on the specific tissue and organ type. For Arabidopsis roots/flowers/leaves, as well as roots of tobacco seedlings, staining can be done without any pretreatment; however, stems and leaves of Nicotiana and P. ) and then dyed. However, for larger tissue samples, vacuum infiltration is required to facilitate the infiltration of cells during the staining session.
(2) Dyeing: Add an appropriate volume of the ready-made dyeing working solution, subject to complete coverage of the material. Wrap in aluminum foil and incubate at room temperature for 3 h overnight. Set up both negative and positive control experiments.
(3) Decolorization: Transfer the dyed material to 70% alcohol for 2-3 times until the negative control material turns white. GUS-positive blue spots are very stable and will not be decolorized by alcohol.
(4) Observation: Most of the blue spots positive for GUS staining can be observed with the naked eye. However, some materials may have very fine blue spots and need to be observed under a microscope.
Note: The blue spots on the white background are GUS expression sites.