BME / DTT Immunoblot

aaa aaa at yahoo.com
Sun Apr 29 07:59:06 EST 2001


In article <200104260502.f3Q52E818094 at ibms.sinica.edu.tw>,
 wolfsc at ibms.sinica.edu.tw ("Wolfgang Schechinger") wrote:

> DTT may be favored aginst BME in your case since the two SH 
> groups present will more likely react with themselves when they 
> become  oxidized at a high dilution (as the conditions in 
> western blot will be). BME should be more likely to react with 
> other molecules (of BME, of antibody or of protein, whatever is 
> present). 

I am not completely understanding of this explinantion.  In my case, BME 
is a poor reducing agent and for reasons unknown to me (but I 
speculate), when I prepare my samples in BME, I loose signal,  and I 
mean a lot of signal (at least factor of 10).  A freind of mine who is  
a protein chemist told me something about BME that may be relevant.  
Most of the preperations of BME that are purchased are not very clean 
and may contain other chemicals that will covalently attach to your 
protein. 
I suspect, The rate of disassociation of BME with a protein is more 
likely to occur than the disassociation of DTT from a protein.  I know 
that DTT is considered to be a more potent reducing agent than BME and 
it works at room temperature.

regards,




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