Hi, what is MW of you want to detect protein? 4~8% gels are used for 15~200 MW protein. For increase efficient to detect high MW protein, need to add 0.01%~0.1% SDS to the transfer buffer. I like use PVDF membrane. If Nitrocellulose membrane maybe change a little. According to the MW of protein, conditions will need to change a little.

Check this paper. SDS-PAGE Electrophoresis of proteins

Writer: Freeman Category Jan. 24, 2010

Good luck

well this system is native gels. sds is not to be used here and we dont do transfer.. the gel as such is put in testing buffer with required substrate and after some time its viewed under UV light n it give flourescnce signal. I am trying to spearate proteasome on the gels, which are high molecular weight protein comles and we dont want to denature them by SDS to maintain the activity of complex.

the complexes are about 750 kD to 2000 kD

Oh, I am so sorry. I haven't experience in this kind of big size complexes.

cloning of Taq DNA gene from thermus aquaticus and in house purification of Taq DNA polymerase. please if someone has experience in this topic specially full thesis and protocol which is easier to get this enzyme.

best regards.

cloning of Taq DNA gene from thermus aquaticus and in house purification of Taq DNA polymerase. please if someone has experience in this topic specially full thesis and protocol which is easier to get this enzyme.

best regards.

this may help you

for cloning

http://journals.mui.ac.ir/rps/article/viewFile/1209/446

http://nar.oxfordjournals.org/content/23/21/4518.full.pdf

for purification

http://bkeelab.bsd.uchicago.edu/Taq%20Purification.pdf

http://faculty.salisbury.edu/~flerickson/protocols/TaqPolPurification.2008.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC331521/pdf/nar00069-0202.pdf