Recent content by sotellme

Moonbear, The reason why i stored my protein samples at -20C is because i followed this kit which said that the protein samples should be stored at -20C. http://www.mrcgene.com/tri.htm Thanks for your awesome tip. I will put them at -70C. :smile:

Dear Moonbear! You saved my life! :smile: actually this is the idea of my advisor. He told me to use different dilution factors. I need to have a talk with him. Thank you! :approve:

I am looking for some ideas about this one. I use B actin as the normalizator in my western blot. i use different dilution factors of the B actin antibody. For some samples i use 500X and for others i use 1000X. I wonder when i normalize my samples and calculate the relative expression levels...

Do you have any ideas why my proteins have less concentrations now than before? Can this liquid form degrade them? Would it be harmful if they are still in this form or should they be better in the frozen form and stored at -70C?

Dear folks, Please help me out. I store my purified protein samples at -20C and they are still at the liquid form (a few are frozen) at this temperature! :cry: I used Trizol kit to purify them and dissolved them in a solution of 9M urea, 4% CHAPS and 30mM Tris/HCl pH 8.5 buffer. They had...

hI iAN, I am thinking of the solvent (?) to dissolve these stuffs. For example for SDS solution, should i use sterile water or buffers?

I have searched and searched till my eyes fall out, but can not find any recipes for making these solutions. Anyone can help me out? 1mM tributylphosphine 1%SDS 10 M Urea

should we add a reducing agent (DTT or 2-Mercaptoethanol) or not in the SDS_PAGE's sample buffer? The plan is using this gel for Western blot.

Can anyone please help me to find the sizes (Da) for these human pituitary proteins? i have looked for them in Entrez and other bioinformatic sites, but am really not sure if they are right. Please, recheck them for me. Thanks a bunch! 1. Glycoprotein hormones alpha chain i have a vERY...

Well, i will work with solid tissue not cells. :confused:

Hey! It sounds like you are familiar with this method. :biggrin: I also plan doing it after running SDs-PAGe. i am going to measure the concentration of each band and for doing it i will use B-actin as a normalizer. My question; how can i detect both the target band and B-actin band in the...

I plan running SDS-PAGE to get the protein profile of human tissues. Which kit is best for this purpose and how can i measure the concentration of a mixture of different proteins? I can measure the protein concentration when it is just a single kind of protein in the mixture, but how about when...

What is the difference between Primers that span introns and primers that flank intron-exon border? Spanning intron does it mean that it covers the whole intron segment? Flanking the border does it also mean that it covers the whole border? :devil: Any ideas would help a lot! Thanks.

Do i need to work on ice when i do Hot start PCR? Hope for inputs. Thanks.

Does the concentration of dNTP in the bottle's label specified the total concentration of the four nucleotides or only each of them? When it says in the protocol "each 200uM" does it mean that i have to add a concentration 4 fold (200uM * 4) of the bottle to get all the four different...