Problem with SDS-PAGE gel - (Aug/11/2013 )

mdfenko on Tue Aug 13 23:50:03 2013 said:

are these the same samples as you've run in the past?

 

if not, is there salt with the sample?

 

are any of the solutions you are using newly prepared (just before your gels went south)?

 

how old is the sds?

Thanks for reply. 

the sample are treated the same way as in the past but the antigen is different. so basically samples are not different.

i dont understand what is meant by salts

We dilute from stock running and transfer buffer

SDS is new.

in what medium is your antigen prior to adding sds sample buffer?

salts such as nacl, kcl, na-acetate, k-acetate, etc.

how old are the stocks.

new lot of sds or newly prepared solution from off-the-shelf lot?

mdfenko on Wed Aug 21 22:23:06 2013 said:

in what medium is your antigen prior to adding sds sample buffer?

 

salts such as nacl, kcl, na-acetate, k-acetate, etc.

 

how old are the stocks.

 

new lot of sds or newly prepared solution from off-the-shelf lot?

I used RIPA lysis buffer and proteins are in it.

yes we use NaCl in RIPA

stocks are fresh 

yes i used newly prepared sds.

newly prepared from off the shelf lot of sds? your sds may be decomposing.

also detergents in the ripa can affect sds binding, salts in ripa can affect migration and heating of the gel.

you may want to confirm that the new stocks are prepared correctly (not intended as an insult, sometimes stuff happens).