Hey,
So I have been working on a project trying to detect whether we have antibodies against a certain protein present in our serum. I have been getting some good results however I'm not sure what the optimal conditions for performing a publish quality stain. I have been using serum at dilutions of 1:100 and 1:10. With a dilution at 1:10 I get a very strong clear signal all the way down to 50ng of protein and a clear signal when the protein is added in with cell lysate. When I use a dilution of 1:100 I get a clear signal down to about 500ng of protein and a signal in cell lysate albeit it is significantly weaker. My control protein that I load gives me a negative signal in both cases.
Does anyone have any advice on what the normal conditions for staining with human serum are as far as both dilution of serum and protein concentration, I'm looking to get a nice clean convincing blot to publish.
Hi Carsten,
Kindly look into the protocol on Western Blot section in the attached research publication by my team. This works really well as I have standardized it myself and repeated it with other molecules too.
All the best
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