Re: 5x sds sample buffer become brown color while preparing
In article , sudheer.pb...@gmail.com says... > > Hello Everyone > > I was trying to prepare 5x sds sample buffer and it has become brown color > instead of blue color. Could anyone please tell me what is the reason and > please tell me your suggestions. > > The 5x sample buffer recipe > > 250mM tris-hcl ph 6.8(1M stock)--10ml > 10% sds-4gr > 50% glycerol(87%stock)--23ml > 0.02% Bromphenol blue-0.008gr > final volume-40ml with DDh20 including 1.4ml of 0.5 > M B-mercaptoethanol but I didnt add here. Promophenolblue is an indicator with a colour chnage from blue to yellow. My guess is that your solution has a pH just around the pKa value of bromophenolblue (3.6). Remember that the pH of most buffers change upon dilution, because it is the activity, rather than the concentation that determines pH. In addition, the high glycerol content may also affect the colour. Is SDS actually soluble under these conditions? -- Car (noun): erratically moving obstacle on the road ___ Methods mailing list Methods@net.bio.net http://www.bio.net/biomail/listinfo/methods
Re: 5x sds sample buffer become brown color while preparing
Sudheer Sangeetham wrote: >I was trying to prepare 5x sds sample buffer and it has become brown color >instead of blue color. Two possibilities: Bromophenol blue is virtually insoluble in water. If the concentration is too high, it stays reddish brown. Try to dilute it with water to 1x and it should turn blue. Then it can be a effect of the pH, since bromophenol blue is a pH indicator as well. Christian -- X-no-Sig: yes ___ Methods mailing list Methods@net.bio.net http://www.bio.net/biomail/listinfo/methods
5x sds sample buffer become brown color while preparing
Hello Everyone I was trying to prepare 5x sds sample buffer and it has become brown color instead of blue color. Could anyone please tell me what is the reason and please tell me your suggestions. The 5x sample buffer recipe 250mM tris-hcl ph 6.8(1M stock)--10ml 10% sds-4gr 50% glycerol(87%stock)--23ml 0.02% Bromphenol blue-0.008gr final volume-40ml with DDh20 including 1.4ml of 0.5 M B-mercaptoethanol but I didnt add here. Thanking you in advance -- Sudheer Babu.S Research Fellow Institute of Biochemistry Biological Research Center Szeged,Hungary. ___ Methods mailing list Methods@net.bio.net http://www.bio.net/biomail/listinfo/methods
