How do you stain Coomassie?
Stain: Dissolve 0.4g of Coomassie blue R350 in 200 mL of 40% (v/v) methanol in water with stirring as needed. Filter the solution to remove any insoluble material. Add 200mL of 20% (v/v) acetic acid in water. The final concentration is 0.1% (w/v) Coomassie blue R350, 20% (v/v) methanol, and 10% (v/v) acetic acid.
Can you leave Coomassie stain overnight?
Note: Gel can be stained for up to 3 hours, but after 3 hours, sensitivity will decrease. If you need to leave the gel overnight in the stain, add 2 ml of 20% NaCl (w/v) in water for every 20 ml of stain. This procedure will not affect sensitivity. Wash the mini-gel with 100 ml of water for 1-3 hours.
How do you Destain instant blue?
Protocol for Gel Drying : Processing the gel prior to full incorporation of stain may result in destaining and reduced sensitivity, this can be reversed by incubating gel in InstantBlue overnight. 2) Submerse the gel in approximately 100 ml ultrapure water at ~70°C (heat for 30s to 60s in a microwave oven).
How do you stain a PVDF membrane after transfer?
Place blot transfer membrane in a plastic box and wash with water three times for 5 min each. Stain membrane with Ponceau S stain for 30 sec to 1 min. Destain membrane with several changes of water for 30 sec to 1 min each, then air dry.
How do you make Coomassie Blue stain?
To make the Coomassie Blue G-250 staining reagent, dissolve 0.2g dye in 100 ml H2O (this will require warming to approximately 50°C). Cool and add 100 ml 2N H2S04. Incubate at room temperature 3 hours to overnight, then filter.
Can you Restain a Coomassie gel?
Yes, you can restain coommassie gels. Just wash the gel after the destain to remove any destain remaining and put the gel back into coommassie stain. Then destain as usual. You can stain and destain with coomassie as many times as you wish.
How does Coomassie blue work?
The Coomassie Brilliant Blue dye which is commonly used in SDS-PAGE, was first described by the German scientist Volker Neuhoff. In the staining reaction, the Coomassie dye binds to proteins through ionic interactions between sulfonic acid groups and positive protein amine groups through Van der Waals attractions.
Can you Coomassie stain a membrane?
Coomassie Blue R250 permanently stains membrane-bound proteins and is compatible with PVDF and nitrocellulose membranes, but it is incompatible with nylon membranes. This technique is relatively insensitive, with a detection limit of ~1.5 μg of protein.
How do you make a Coomassie stain?
To prepare 1 l of a staining solution, dissolve 1 g of Coomassie R250 to 300 ml of methanol. Then add 650 ml of MQ water and 50 ml of acetic acid. Stir the solution on a magnetic stirrer for 2 h. The solution can be filtered through a Whatman No.
What does Coomassie Blue stain tell you?
Coomassie Blue stain is used to stain the protein bands in polyacrylamide gels. One common way to use it is to dissolve the dye in a mixture of methanol, acetic acid, and water. This stain will permeate the gel, stain the protein, and also fix the protein in place.
What is the Abcam product code for Coomassie?
ab119211InstantBlue®Coomassie Protein Stain product of Expedeon, an Abcam company Applicable to Expedeon product codes: ISB1L For the rapid Coomassie blue staining of protein in polyacrylamide gels.
What do I need for Coomassie blue staining?
You may use any Coomassie™ staining protocol of choice. You will need following items for Coomassie Blue Staining. 0.1% Coomassie® R-250 in 40% ethanol and 10% acetic acid (if using Invitrogen™ Coomassie™ R-250 Staining) Destaining Solution consisting of 10% ethanol and 7.5% acetic acid (if using Invitrogen™ Coomassie™ R-250 Staining)
How to use Coomassie stain after SDS-PAGE?
Only use the Coomassie stain on gels post-transfer to check the efficiency of the transfer, or if you have no plans to transfer and just want to observe the results of the SDS-PAGE separation. As soon as the power is turned off the separated protein bands will begin to diffuse (they are freely soluble in aqueous solution).
Which Coomassie staining protocol should I use for Invitrogen™ Novex™ gels?
The coomassie staining protocol described below is recommended for staining Invitrogen™ Novex™ Gels. You may use any Coomassie™ staining protocol of choice. You will need following items for Coomassie Blue Staining.