Can 2-D lysis buffer be used for western Blotting?

Does anyone know what might be causing the smeared bands on my western blot?

I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...

10 August 2024 7,477 3 View

What will be the best way to test NFkb activation via western blot?

Hi, I have lysate from human macrophages, and I wonder what will be the best way to check the NFKB activation in those cells. I was thinking p-p65 or p-ikkb./ Is that make sense?

18 July 2024 5,214 4 View

Trouble finding an anti-Digoxigenin antibody to perform Southern Blot technique?

I'm currently using Roche anti-DIG-AKP Fab fragments to perform Southern Blot assays, but this reagent is delayed since november last year. It's anybody facing the same problem? If so, how are you...

15 July 2024 6,667 0 View

How to detect phosphorylation on Western Blots?

Having trouble detecting phosphorylated HSL. Any tips for how to prevent de-phosphorylation events or detect phosphorylation?

15 July 2024 2,054 3 View

Is there anyway to distinguish two proteins of same molecular weight?

I know mass spectrometry will work. But, is there any simple and straightforward method to distinguish between the two proteins of the same molecular weight? They are cryptochromes and...

14 July 2024 9,322 2 View

What would you do next?

Studying a specific monogenetic disease based on mutations in different individual genes. I try to recreate the disease phenotypes of donors in vitro. I knocked out a gene x with above 90%...

12 July 2024 6,312 0 View

Loading control for western blot to detect Histone H1.2?

Hello, I isolated total histones successfully from the skin primary cells by acid extraction. I wanted to detect histone H1.2 expression in different conditions by western blot. Can some one...

10 July 2024 9,164 1 View

Personal recommendations for GAPDH antibody?

Hello! I am planning a western blot experiment, and I am looking for GAPDH antibodies to order. In the past, I have experienced a lot of hit-or-miss when ordering antibodies based on just the...

07 July 2024 1,683 0 View

How can I solve this problem with my western blot?

I have this problem with my western blot for the ABCG2 antibody on nitrocellulose membrane. Help please

02 July 2024 8,713 2 View

How to troubleshoot error E1 on Biorad Powerpac Basic despite using sufficient and fresh buffer?

I have used this BIorad Powerpac Basic for 4 years now, before which it was used for another 5-6 years. Generally we addressed the E1 error by compensating with fresh buffer and/or filling the...

01 July 2024 7,104 1 View

Lichuan Hong

I have a same question,after my 2D electrophoresis /MALDI TOF-TOF identified some biomarker,i need to validate them.For saving lysis tissue sample,i try to use them to do western blot,but i find some problem during western blotting.

Ravi Shankar Singh

Thank you Mr Lichuan Hong. what type of problem did you face?