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FAQ - AcquaStain Protein Gel Stain


Questions

Answers

Q1
Why is it better to use AcquaStain at room temperature?   A1 We recommend allowing AcquaStain to equilibrate to room temperature before staining. AcquaStain works much more quickly at warmer temperatures. In fact it can take 10x longer to visualize bands using cold AcquStain. Read A2/Q2 for more detail.
Q2
What is the optimal temperature for staining gels?   A2 For achieving the most effective staining, we recommend preheating the appropriate amount of AcquaStain (25ml for a mini-gel) to a temperature between 37°C and 42°C.
Q3
What does Acqua Stain bind to?   A3 Acqua Stain binds to basic amino acids in a similar way standard coomassie does, only much faster and more sensitive.
Q4
Can I further decrease the time for staining?   A4 Yes. By placing the gel in Acqua Stain and microwaving for 10-20 seconds, you can cut the times by roughly 50%.
Q5
Can I increase the sensitivity/contrast on my Acqua Stain Gels?   A5 Yes. If you gently agitate the gel in stain at 50°C during staining, you will see increased sensitivity. If you gently agitate the gel in heated deionized water (50-60°C) for 15-25 minutes post staining, you will see increased contrast.
Q6
Why do the protein bands on my gel appear as blobs when I incubate overnight?   A6 It is not possible to overstain however, Acqua Stain is more sensitive than traditional methods so it is quite possible that the gel was overloaded. In addition, low molecular weight proteins can diffuse over time if the gel isn't fixed. We recommend decreasing the stain time to 60 minutes or less or loading less onto the gel.
Q7
Does Acqua Stain fix the gel?   A7 No. Acqua Stain is an aqueous-based stain and therefore will not fix your gel. After 30 minutes of incubation with the stain, the gels are very stable so there is no need to fix the proteins.
Q8
After staining, can I store the gel in acetic acid?   A8 Yes. You can store the gel in acetic acid or deionized water.
Q9
Can Acqua Stain be used in colorimetric assays?   A9 Yes. You can measure with a standard spectrophotometer or plate reader at 595nm
Q10
Can proteins stained with Acqua Stain be used in mass spec analysis?   A10 Yes. You can see the protocol here.
Q11
What is the protocol for dyring down the gels after staining?   A11 Yes. You can see the protocol here.
Q12
Is AcquaStain compatible with a western blot?   A12 Yes. Because it reversibly binds, you can use it for a western blot. You will need to wash the gel a few times in transfer buffer. You can also stain the gel after blotting as this is a good method of determining transfer efficiency.
Q13
Can AcquaStain be reused?   A13 Yes. Typically, customers can use it two or three times.