Search found 20 matches

by samkh918
Tue Apr 13, 2010 11:10 pm
Forum: Molecular Biology
Topic: Loading control for different human tissue samples
Replies: 1
Views: 2672

Loading control for different human tissue samples

Hello, I have ordered an immunoblot (NC membrane) for western blotting, which contains different human tissues such as kidney, spleen, prostate, etc. in each lane. I want to see if equal amount of proteins have been loaded in each lane as promised by the company but my problem is that I can't use B-...
by samkh918
Sat Nov 28, 2009 9:36 pm
Forum: Molecular Biology
Topic: Silver staining after coomassie blue
Replies: 2
Views: 6343

Re: Silver staining after coomassie blue

Thanks a lot!
by samkh918
Mon Nov 09, 2009 7:53 pm
Forum: Molecular Biology
Topic: Silver staining after coomassie blue
Replies: 2
Views: 6343

Silver staining after coomassie blue

Hello, I stain my 2D gels with coomassie blue and sometimes about a week later, perform silver staining on them. I was wondering if the original coomassie blue staining has any effect on the quality of the subsequent silver staining. I would also very appreciate if someone could send me a tested sil...
by samkh918
Mon Nov 09, 2009 7:42 pm
Forum: Molecular Biology
Topic: pH marker for 2D electrophoresis
Replies: 0
Views: 1190

pH marker for 2D electrophoresis

Hello, I use 7cm, pH4-7 IPG strips in my 2D electrophoresis experiments. After the second dimension run and staining the gel with coomassie blue, I see the spots and I want to know at what pH some spots are located. I was wondering if there is a way or a pH marker that can be used to find out the ex...
by samkh918
Sun Sep 20, 2009 8:52 pm
Forum: Molecular Biology
Topic: Matching 2D membrane spots to the gel
Replies: 1
Views: 2558

Matching 2D membrane spots to the gel

Hello, I am performing two-dimensional (2D) electrophoresis experiments and for each sample I make two IPG strips. After performing the first dimension for both strips, I stain one gel with coomassie blue to see all the spots, and use the other gel to transfer the proteins to a nitrocellulose membra...
by samkh918
Wed Aug 12, 2009 1:56 pm
Forum: Cell Biology
Topic: Antibiotic effect
Replies: 5
Views: 14541

Re: Antibiotic effect

Thanks a lot for your feedback. I have another question along the same line. Is it common for bacteria in a cell tissue culture to enter the cells or are they mainly present in the culture media? If they enter the cells would the added antibiotic be still effective?

Thanks for your help.
by samkh918
Tue Aug 11, 2009 5:38 pm
Forum: Cell Biology
Topic: Antibiotic effect
Replies: 5
Views: 14541

Antibiotic effect

Hello, Is it true that antibiotics such as streptomycin can only suppress the growth of bacteria in a cell culture dish and if there were already bacteria in the dish, once you stop adding the antibiotic, they will start to grow? Are there any antibiotics that actually kill the bacteria so that you ...
by samkh918
Tue Apr 14, 2009 6:23 pm
Forum: Molecular Biology
Topic: Cell lysis and Lowry assay for 2D electrophoresis
Replies: 0
Views: 2145

Cell lysis and Lowry assay for 2D electrophoresis

Hello, I want to perform a 2D electrophoresis on my neuronal cells and wanted to get your feedback on some of my questions: 1)Can I use my 2D sample buffer: 8M Urea, 2% w/v CHAPS 50mM DTT 0.2% w/v100X Bio-Lyte ampholyte 0.002% w/v BPB for the cell lysis step as well? I am going to try both sonicatio...
by samkh918
Fri Dec 12, 2008 2:48 am
Forum: Cell Biology
Topic: Chaperone activity assay
Replies: 1
Views: 2554

Chaperone activity assay

Hello,

I was wondering if someone could direct me to one or more chaperone activity assay, other than the luciferace assay which I am aware of. I want to use this assay for heat shock proteins, Hsps.

Thank you in advance for your help.
by samkh918
Thu Aug 14, 2008 7:38 pm
Forum: Molecular Biology
Topic: Gel extraction kit
Replies: 1
Views: 1807

Gel extraction kit

Hello, I was wondering if someone could explain the main purpose of the gel extraction kits. In some protocols I see that after performing PCR on the vector containing gene of interest, the amplicon is run on the gel first and the extracted using gel extraction kits. They then proceed with A-additio...
by samkh918
Thu Aug 14, 2008 7:34 pm
Forum: Molecular Biology
Topic: Glycine in Western Blot buffers
Replies: 2
Views: 19466

Re: Glycine in Western Blot buffers

Thank you for your explanation.
by samkh918
Tue Aug 12, 2008 12:51 am
Forum: Molecular Biology
Topic: Glycine in Western Blot buffers
Replies: 2
Views: 19466

Glycine in Western Blot buffers

Hello,

Could someone please let me know what's the reason for adding glycine to the Western blot running and transfer buffer?

Thanks a lot.