Decoloration-free polyacrylamide gel protein rapid staining solution and preparation and use methods thereof
A polyacrylamide gel protein and rapid staining technology, applied in the field of protein analysis and detection, can solve the problems of low sensitivity, long process time, cumbersome operation, etc., achieve high sensitivity, simple steps, and improve work efficiency
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Embodiment 1
[0041] Example 1: Effect of Adding Ammonium Sulfate and Cyclodextrin on Protein Staining in Coomassie Brilliant Blue Staining Solution
[0042] The configuration of the protein staining liquid of the present embodiment: the ratio of each component of each group is as follows
[0043]
[0044] With 12.5% 1.0mm thickness SDS-PAGE gel, 150V electrophoresis for 45min. The protein samples are pre-stained protein marker, non-pre-stained protein marker, BSA 2000ng 1000ng, 500ng, 250ng, 125ng, 62.5ng, 30ng, 20ng, 15ng, 10ng, 5ng, bacterial lysate.
[0045] After electrophoresis, put the gel into an appropriate amount of prepared protein staining solution, stain for 5 minutes, and take it out for observation and analysis after 60 minutes.
[0046] figure 1 Pictures were taken after 5 min of staining. Group a and group c can only stain more than 1000ng of protein bands, group b and group d can stain more than 30ng of protein bands, so it can be analyzed that a-cyclodextrin can e...
specific Embodiment 2
[0048] Specific embodiment 2: the impact of Coomassie Brilliant Blue concentration on dyeing
[0049] The protein staining solution provided in this example includes the following components: absolute ethanol 15% (v / v), ammonium sulfate 4% (w / v), and 1.5% β-cyclodextrin (w / v). Coomassie Brilliant Blue G-250 concentrations were: group a 0.01% (w / v); group b 0.015% (w / v); group c 0.025% (w / v);
[0050] In this implementation, the electrophoresis conditions and protein samples are the same as those in implementation 1.
[0051] After electrophoresis, put the gel into an appropriate amount of prepared protein staining solution, stain for 5 minutes, 15 minutes, 30 minutes, and 60 minutes respectively, and observe and analyze.
[0052] The results showed that as the staining time increased, the stained bands became darker. For the same staining time, a higher concentration of Coomassie Brilliant Blue staining solution will make the protein bands stain darker, but no significant incr...
Embodiment 3
[0053] Embodiment 3: ammonium sulfate concentration optimization
[0054] The protein staining solution provided in this example includes the following components: absolute ethanol 15% (v / v), G-250 0.015% (w / v), and 1.5% β-cyclodextrin (w / v). Ammonium sulfate concentrations were: group a 2% (w / v); group b 4% (w / v); group c 6% (w / v); group d 8% (w / v).
[0055] In this implementation, the electrophoresis conditions and protein samples are the same as those in implementation 1.
[0056] After electrophoresis, put the gel into an appropriate amount of prepared protein staining solution, stain for 5 minutes, 15 minutes, 30 minutes, and 60 minutes respectively, overnight, and take it out for observation and analysis.
[0057] The results showed that the staining background decreased with increasing ammonium sulfate concentration. like Figure 4 Shown: The pictures taken after staining for 30 minutes showed no significant difference in the staining background. The staining time i...
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