Alternative methods for western blotting?

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Non-specific bands in PCR?

Usually I am doing PCR with 0.5 ul of Taq polymerase (abm) per reaction. Since I wanted to increase the sensitivity of target amplification, I tried PCR with 1.0 ul of Taq. It gave some bands in...

02 March 2018 9,197 6 View

Mass spectroscopic analysis for protein identification?

I am doing immunoprecipitation for concentrating by target proteins. I usually elute my proteins in SDS loading buffer containing Tris, glycerol, SDS, beta mercaptoethanol and bromophenol blue....

11 December 2017 1,246 2 View

Human Anti-Leishmania IgG ELISA Kits...

I have found two commercial ELISA kits for detection of human leishmaniasis. 1. Human Anti-Leishmania IgG ELISA Kit (ab108749) by Abcam 2. Leishmania IgG ELISA Kit (KA3295) by Abnova Both are...

03 April 2017 3,701 1 View

Why did the protein sample and marker not migrate properly in the separating gel (SDS-PAGE)?

I am facing the a problem in my SDS page gel. protein marker and samples not get well separated. All samples were stop separating at the middle of separating gel (protein marker also). Dye front...

09 October 2016 3,411 12 View

Physical characterization of crude parasite Ag???

Is there a simple method to physically characterize the crude Ag extracted from a parasite to check whether they are proteins, polysaccharides or phospholipids like compounds? I am using crude Ag...

07 August 2016 4,034 3 View

Analysis of proteins using westernblotting?

Does 3D structure of a protein affect for migrating distance of a protein?

06 July 2016 6,313 0 View

What is the best condition for blocking western blot membrane?

Can I keep my blocking buffer (skimmed milk or BSA) at room temperature for overnight? Is that affect for my protein?

05 June 2016 3,976 5 View

Commercial protein sequencing services?

I want to sequence some unknown protein excised from 1D-SDS-gel. Have anybody experienced with protein sequencing? Are there any good commercial protein sequencing services?

04 May 2016 4,567 1 View

Fuzzy bands on SDS-PAGE?

What are the reasons for getting fuzzy bands on SDS-polyacrylamide gel?

03 April 2016 594 9 View

TMB SUBSTRATE FOR WESTERN BLOT???

How can we convert TMB of ELISA for western blot by converting soluble TMB product of ELISA into insoluble with addition of 0.05% sodium nitroprusside? Can I use ELISA TMB substrate for Western blot?

02 March 2016 8,684 0 View

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 7,480 3 View

Do I need an antigen retrieval for a multiplex RNAscope experiment for adherent cells?

Hello! I planned to do an multiplex RNAscope fluorescent assay for cultured cells in a 96 well plate format according to the protocol provided by ACD:...

22 July 2024 3,334 0 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

ELISA sandwich with two monoclonal antibodies ?

hi, I am facing difficulties in the développement of an ELISA sandwich. I have several experiences in developing ELISA but that’s the first time I can’t find any solution. My test is currently as...

16 July 2024 9,044 10 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 2,055 3 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 6,668 0 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 9,324 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 6,313 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 9,165 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 1,685 0 View