The main purpose
YIJI protein silver nitrate staining reagent is a kind of silver nitrate gel which is designed to be directly fixed on the polyacrylamide gel by fixing, washing and dyeing with silver nitrate.
Color, the authoritative and classic technical method of producing silver bands with a sensitivity of 10 nanograms or less on a clear background. Master of Science
The scientist has carefully developed and successfully tested it. It is suitable for protein electrophoresis including detection of IEF. The product is ready to use, the performance is stable, and the operation is easy.
It has high sensitivity, clear color and good repeatability.
technical background
The silver ion of silver nitrate (AgNO3) combines with the protein to chemically reduce it to visible metallic silver, showing a dyeing effect. Usually used for protein
Detection of polyacrylamide gel electrophoresis. Compatible with mass spectrometry.
product content
YIJI Fixative (Reagent A) ml
YIJI treatment solution (Reagent B) ml
YIJI Purifying Liquid (Reagent C)
YIJI staining solution (Reagent D) ml
YIJI Color Reagent A (Reagent E) ML
YIJI Color Reagent B (Reagent F) Microliter
YIJI Stop Solution (Reagent G) ML
YIJI Decolorizer A (Reagent H) ML
YIJI Decolorization Solution B (Reagent I) ML
Product manual 1 copy
storage method
Save YIJI staining solution (Reagent D) in a 4°C refrigerator to avoid light, and keep the rest at room temperature for 12 months.
User-supplied
Plastic tray: container for dyeing
Ion-free water: used to dilute the kit solution
Constant speed oscillator: for incubation during dyeing operations
Experimental procedure
Before the start of the experiment, complete the SDS-PAGE protein electrophoresis run, take out the colloid, make the orientation mark, and then do the following:
First, the fixation process
1
1. Prepare a milliliter of beaker and make it to the number mark
2. Extract ML YIJI Fixative (Reagent A) into the beaker, add ML of non-ionized water, and mix.
3. Add to the size of the dye tray
4. Put the polyacrylamide gel into the dyeing tray
5. Incubate for a few minutes on a constant speed (RPM) shaker at room temperature,
6. Simultaneously extract ML YIJI treatment solution (Reagent B) into the beaker, add ml of non-ionized water, and mix.
7. Carefully pour off the YIJI Fixative in the staining tray (Reagent A)
8. Add the YIJI treatment solution (Reagent B) diluted in the milliliter beaker.
9. Incubate for minutes on a constant speed (RPM) oscillator at room temperature
10. Carefully pour off the YIJI treatment solution (Reagent B) in the dye tray
11. Repeat the experiment steps twice
12. Simultaneously extract ML YIJI Purifying Fluid (Reagent C) into the beaker, add ML of non-ionized water, and mix.
13. Add the YIJI Purifying Solution (Reagent C) diluted in the milliliter beaker to the dyeing tray.
14. Incubate for minutes on a constant speed (RPM) oscillator at room temperature
15. Carefully pour off the YIJI Purifying Solution in the dye tray (Reagent C)
16. Repeat the experiment step once
Second, dyeing treatment
17. Direct extraction of milliliter YIJI staining solution (Reagent D) , added to the dyeing tray
18. Incubate for minutes on a constant speed (RPM) oscillator at room temperature
19. Carefully pour off the YIJI staining solution (Reagent D) in the staining tray
20. Add milliliters of self-contained, ion-free water
twenty one. Incubate for minutes on a constant speed (RPM) oscillator at room temperature
twenty two. Carefully pour off the ion-free water in the dye tray
twenty three. Repeat the experiment step once
Third, color processing
twenty four. Simultaneously extract milliliters of ion-free water to the beaker
25. Add microliter YIJI Reagent E and Microliter YIJI Reagent F to mix
26. Add YIJI color working solution in the milliliter beaker to the dyeing tray
27. Incubate to a constant temperature (RPM) shaker at room temperature for a minute, and the protein silver band begins to appear (note that the color is not too deep)
28. Carefully pour off the YIJI coloring solution in the staining tray (if the staining is too deep, see step 30)
29. Add ML YIJI Stop Solution (Reagent G) directly until all photos and analysis are completed
Fourth, bleaching treatment
30. If the staining is too deep, remove ML YIJI Decolorizer A (Reagent H) and ML YIJI Decolorizer B (Reagent I)
To the beaker, add ML of non-ionized water and mix
31. Add YIJI bleaching work solution to the dyeing tray.
32. Incubate at room temperature on a constant speed (RPM) shaker until the strip color returns to a satisfactory state
33. Carefully pour off the YIJI bleaching solution in the dye tray
34. Add in milliliters of ion-free water for cleaning
Five, mass spectrometry pretreatment
2
35. If mass spectrometry is required, remove microliters of YIJI Decolorizer A (Reagent H) and microliters of YIJI Decolorizer
B (Reagent I) to the milliliter centrifuge tube, add microliter of non-ionized water, mix
36. Carefully cut the protein band gel
37. Put in the milliliter centrifuge tube until the strip is completely discolored
Precautions
1. This product is 10 X 25 ml specifications
2. Each 25 ml kit solution is used to treat 6cm X 8cm size colloids; every 50ml kit solution is used to process 12cm X 12cm large
Small colloid; the larger the colloid, the increased the amount
3. Wear gloves when handling
4. The reagent solution must be diluted with ion-free water.
5. Incubation temperature is room temperature
6. The company provides 3% (YJ12034.1), 8% (YJ12034.3), 10% (YJ12034.4) SDS-PAGE solution and protein Coomass
Bright Blue Staining Kit (YJ30003), YIJI Protein Nile Red Staining Kit (YJ30007), YIJI Protein SYPRO Orange
Color staining kit (YJ30008)
Quality Standard
1. This product has been certified to be stable.
2. This product has been proven to have excellent dyeing effect.
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