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Bubble test - (Feb/19/2009 )

Does some body knows what is the bubble test to locate the protein in the series of the fraction.
I shall be very thankful to them
regards

-samita-

Hey

What are you doing? Purifying protein and want to know which fraction has yr protein?

You can take abt 20 ul of the fraction and see if it turns bradford blue. Its a quick indication whether yr protein is there or not.

Best
TC

-T C-

Can you tell me little bit more about it how to do it......

T C on Feb 19 2009, 07:04 AM said:

Hey

What are you doing? Purifying protein and want to know which fraction has yr protein?

You can take abt 20 ul of the fraction and see if it turns bradford blue. Its a quick indication whether yr protein is there or not.

Best
TC

-samita-

Hey

I missed yr post....so a late reply.

I use the standard bradford that we get from biorad and dilute it 1:5. Take 100 ul of this solution in a 96 well plate and add 20ul of protein to check for blue colour, indicating presence of protein.

Best
TC

-T C-

samita on Feb 19 2009, 12:33 PM said:

Does some body knows what is the bubble test to locate the protein in the series of the fraction.
I shall be very thankful to them
regards


Hi
I think that what is meant by the bubble test is that if you flick the fraction tubes frothy bubbles will appear if high concentrations of protein are there. It's not a quantitative test like the Bradford assay but just a quick way to decide which fractions to look at more closely

P

-Penguin-

Hi Penguin
You are absolutely right... I aksed about this becasue I have NADH in my buffer and it gives a high absorbance on UV and I could not trace out which fraction contains my protein. If you can explain me little bit more then its a good help of me.
How much fraction volume is required for this.
How quickly or fast flickering one have to do....
Its normal t have bubble in the solution if you flick the solution but what makes a differnece between the rotein fraction or the normal fraction.
I shall be very thankful to you for your kind reply.
best regards

Penguin on Feb 24 2009, 04:30 AM said:

samita on Feb 19 2009, 12:33 PM said:

Does some body knows what is the bubble test to locate the protein in the series of the fraction.
I shall be very thankful to them
regards


Hi
I think that what is meant by the bubble test is that if you flick the fraction tubes frothy bubbles will appear if high concentrations of protein are there. It's not a quantitative test like the Bradford assay but just a quick way to decide which fractions to look at more closely

P

-samita-

samita on Mar 4 2009, 09:57 AM said:

Its normal to have bubble in the solution if you flick the solution but what makes a difference between the protein fraction or the normal fraction.

the bubbles will be persistent in the fractions containing protein.

try it and you will see.

-mdfenko-

Hi,

Give the tube 3 or 4 hard flicks with your finger, bubbles should appear that don't go away quickly (kinda like the head on a beer!!)

How many fractions do you have? Is it too many to pick out every other one and run it on SDS-PAGE/test by Bradford assay?

P

-Penguin-