GUS Staining Kit (Ready-to-use)

Cat. No.: G1508328

GRADE & PURITY BioReagent, Biological Stain, for Microscopy

Overview

GUS Staining Kit (Ready-to-Use) employs a classic histochemical method to rapidly detect gus gene fusion markers in plants through a color reaction with a substrate.

The gus gene is present in the genomes of certain bacteria such as Escherichia coli. Its product, β-glucuronidase (GUS), is a hydrolytic enzyme that catalyzes the hydrolysis of many β-glucuronide esters. Because most plant cells lack endogenous β-glucuronidase background activity, the gus gene is one of the most commonly used reporter genes in transgenic plants, especially widely applied in studies of transient expression of foreign genes. β-Glucuronidase can cleave 5-bromo-4-chloro-3-indolyl-β-D-glucuronide (X-Gluc) into an insoluble blue product. Therefore, when transgenic plants carrying the gus gene are soaked in a staining solution containing X-Gluc substrate, tissues or sites with GUS activity appear blue or show blue spots, which can be observed with the naked eye or under a microscope. The detection method is simple, fast, sensitive, stable, and has low background activity.

This kit contains all reagents required for GUS staining. It is convenient to use: simply mix the prepared X-Gluc Solution and GUS Buffer in the specified ratio to prepare the GUS staining solution.

G1508328	Component	50 mL	100 mL	Storage
G1508328A	X-Gluc	1 EA	2 EA	-20℃. Store in the dark.
G1508328B	X-Gluc Solution	1 mL	1 mL×2	RT. Store in the dark.
G1508328C	GUS Buffer	50 mL	100 mL	2-8℃. Store in the dark.

Reagents, consumables and Equipments not provided

• 70% ethanol, Deionized water

• Small vials or multi-well plates, Microscope

Instructions for Use (For Reference Only)

1. Prepare X-Gluc Solution (50×): Add 1 mL of X-Gluc Solution completely to one vial of X-Gluc. Dissolve thoroughly to obtain the X-Gluc Solution (50×). Store the prepared X-Gluc Solution (50×) at -20°C; it is stable for 1 month.

2. Prepare X-Gluc Staining Solution: Take an appropriate amount of X-Gluc Solution (50×) and GUS Buffer, mix thoroughly at a ratio of 1:49 to prepare the X-Gluc staining solution. For example, add 0.2 mL of X-Gluc Solution (50×) to 9.8 mL of GUS Buffer to prepare 10 mL of GUS staining solution. It is best to prepare the staining solution fresh for each use. For short-term storage, it can be kept at 4°C for up to 3 days.

3. Staining: Add an appropriate amount of tissue (e.g., leaves) to be stained into sufficient GUS staining solution to completely submerge the tissue.

4. Incubation: Incubate at 37°C for 1–24 hours. Blue color gradually appears with extended incubation time. Sites with high GUS activity will show blue or blue spots.

5. Decolorization: Remove chlorophyll from the sample using 70% ethanol. Generally, submerge the sample in ethanol for 1–3 hours until negative controls turn white. If necessary, repeat the decolorization step to thoroughly remove chlorophyll. Store the sample in ethanol. Observe with the naked eye or under a standard light microscope; blue color on a white background indicates GUS expression sites.

Notes

1. It is best to prepare the GUS staining solution on ice and use it immediately. Unused prepared solution can be stored at -20°C protected from light. Discard if the solution turns red or purplish-red.

2. Due to tissue specificity and other factors, the intensity of the blue color reaction may vary. Optimal experimental conditions should be determined empirically.

3. Tissues such as Arabidopsis roots, flowers, leaves, and tobacco seedling roots can be stained directly. Stems and leaves of plants like tobacco and potato should be cut into thin slices (1–3 mm) before staining.

4. For large tissues or samples, vacuum infiltration can be used to facilitate substrate and enzyme penetration into cells.

5. The GUS Staining Kit (Ready-to-Use) offers good staining sensitivity, but non-specific reactions (false positives) may occasionally occur.

6. Use reagents as soon as possible after opening to avoid affecting subsequent experimental results.

Storage and Shipping

Storage

Store at 2-8°C,Protected from light,Room temperature,Store at -20°C

Shipped In

Ice chest + Ice pads

Stability And Storage

Each component has a shelf life of 1 year under corresponding storage conditions.