Silver Stain kit for Protein, , 25T

Product Information

Product Description/Introduction

Silver staining is a highly sensitive staining method, the basic principle of which is to display the protein band by forming a black precipitate on the protein by the reduced anion. The sensitivity of silver stain is 100 times higher than the traditional Coomassie brilliant blue stain and can detect proteins below 0.5 ng. Because of its high sensitivity, silver staining is widely used in 2D gel analysis and vertical PAGE gel for very low protein content determination. It is a common method for the detection of low abundance proteins in gels. This product can be used for protein detection in SDS-PAGE or PAGE polyacrylamide gel, with high sensitivity, easy to operate, almost colorless background, silver staining of one gel can be completed within 90 min. This kit can stain 25 gels of the same specification.

Storage and Shipping Conditions

Ship at room temperature; The Silver Stain Enhancer store at room temperature, and the other reagents store at 2-8, valid for 12 months.

Product Components

Assay Protocol/Procedures

1. Fixation: Prepare fixation solution according to the table below. Put the gel in a clean dye bath, wash three times with ultra-pure water, add 20 mL of fixing solution to cover the gel, and incubate on a horizontal shaker (60-70 rpm) for 30 min. To prevent acetic acid and ethanol volatilization, please seal the dye bath with plastic wrap. Prolonged fixation time can further reduce the background.

2. Washing: Pour out the fixing solution, replace the ultra-pure water covering gel, and shake the horizontal shaker for three times, 10 min each time.

3. Sensitization: Prepare the sensitization working solution according to the table below. Add the sensitization solution to cover the gel and incubate for 1 min on a horizontal shaker with slow shaking.

4. Washing: Pour out the sensitization working solution, replace the ultra-pure water and clean the shaker with three quick shakes of 20 s each time.

5. Silver dyeing: Prepare silver dyeing solution according to the table below. Add the silver staining solution to cover the gel and incubate for 20 min by shaking slowly. Silver dyeing solution should be used quickly within 2 h.

6. Wash with water: Discard the dye solution, replace the ultrapure water and wash the shaker three times with a quick shake for 20 s each time.

7. Color development: Prepare the developing solution according to the table below. Add the developing solution to cover the gel and incubate with slow shaking for 1-5 min until the desired target protein band appeares.

8. Termination: Prepare the development termination solution according to the table below. Add the development termination solution to cover the gel and incubate for 2 min with fast shaking on a shaker. It is normal for CO2 gas to be produced during the process. Then discard the termination solution and wash the gel with ultrapure water for 2-5 min.

9.  Storage: After cleaning, the gel can be placed for observation and photography. The stained gels can be stored in ultrapure water or 1% acetic acid solution, or dried in an appropriate manner.

Note

1. To prevent interference from impurities, use high purity water (resistance > 16 MΩ/cm) and clean glassware or plastic containers. Do not use metal containers.

2.  Please take protective measures throughout the experiment. Ultrapure water, absolute ethanol and glacial acetic acid should be prepared by yourself.

3.  Do not directly touch or press the colloid during the operation, and do not use metal for the dyeing tank, glass or plastic containers are the best.

4. Store the Silver Stain Enhancer at room temperature away from light. Do not store it at 4, otherwise it will fail.

5. All working solutions shall be prepared and used at once. After opening the reagent, tighten the cap to prevent the solution from evaporating and reacting with airborne substances, which will affect the dyeing effect.

6. The developing solution is stored at 4 for a long time with crystal precipitation (normal phenomenon), and can be incubated at 37 until completely dissolved before use.

7. When the gel is thick or the operating temperature is low, the color development time can be extended appropriately.

8. The dyeing time, washing time, color development and termination time of each step should be well controlled, otherwise the background of the gel will become darker and the target bands are not easily observed.

9. For your safety and health, please wear safety glasses, gloves, or protective clothing.

For Research Use Only!

https://www.servicebio.com/goodsdetail?id=7906