Western of His-tagged protein.
I want to perform splenocyte restimulation culture for the estimation of Th1 and Th2 cytokines by ELISA. As each groups consists of 5 mice immunized with a test vaccine preparation; can splenocyte...
08 September 2019 4,812 2 View
My protein (64kDa) expressed in insoluble inclusion bodies. I tried to purify it by ion-exchange and affinity but unable to purify protein. So, want to try ammonium sulfate precipitation of...
02 March 2018 8,364 5 View
I want to amplify a gene whose end terminus sequence is in such a way that the after designing NotI restriction site inserting (reverse) primer; it forms primer dimer during PCR amplification. I...
08 September 2017 7,996 3 View
I am working on recombinant protein (Ig's) expressed in form of inclusion bodies. Upon giving certain treatments there is a up shift in protein band. If my protein is degrading during the process;...
10 November 2016 7,922 4 View
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
Hello everyone, I am currently working on a protein that is 10x his-tagged and positively charged (predicted pI=10.16). But when I tried to use Ni-NTA column to purify the protein, it's not...
24 July 2024 7,293 4 View
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
Good day everyone, Trying to perform protein purification for the first time, gotta buy the whole equipment/reagents. Protein of interest 48 kDa (Antithrombin), expressed from his-tagged plasmid...
16 July 2024 3,692 6 View
Having trouble detecting phosphorylated HSL. Any tips for how to prevent de-phosphorylation events or detect phosphorylation?
15 July 2024 2,055 3 View
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
I am treating human cells with an AAV vector encoding for my gene of interest tagged by FLAG. I want to quantify the mRNA expression of my gene of interest via qPCR to discriminate the endogenous...
14 July 2024 8,482 3 View
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
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
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
