Questions pertaining to western blots, which are used for detecting specific proteins (sometimes called protein immunoblots).

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6
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Optimal pH of protein buffer? Basic principles to adjust buffers according method and analysis

Protein buffers such as PBST, which is used in western blotting, are normally adjusted to pH 7.4. When I try to find why, I find some information about optimal pKa for protein stability. Im not sure I ...
3
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65 views

Relative densitometry from SDS PAGE

I'd like to perform densitometry on a Coomassie stained SDS PAGE gel to compare a recombinant protein's expression levels under two conditions. I'm using BioRad's Image Lab software. My questions are ...
2
votes
0answers
73 views

Why HIS tag is not detected in WB or dot blot?

I would like to know why the recombinant protein with his tag at c terminus is not being detected neither in western blot or dot blot. I'm trying to validate protein expression of four proteins. ...
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0answers
14 views

Using only one antibody to detect COX-2 enzyme

If I wanted to detect COX-2 from a western blot test, would only a primary antibody work? I'm on a budget so does anyone know of relatively cheap COX-2 antibodies?
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18 views

High intensity background on western blot membrane?

I did western blot and I got the high intensity background only in the upper part of the membrane and there is a horizontal line around 50-70kDa (this line separate the upper part and lower part of ...