What is the Best Method of Stripping for Reprobing (WB) Phosphorylated Proteins? e.g Insulin and proliferation pathway proteins.
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
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05 August 2024 4,407 0 View
Hello, I am currently analyzing some phosphoproteomics data, but I have peptides with multiple phosphorylation sites or phosphorylations together with carbamidomethylation or oxidation. How can I...
04 August 2024 8,432 3 View
I am trying to prepare samples for IP. The pellet size is good and I am using 1% Tx-100 and using a syringe to further lyse the cells and maintain protein-protein interactions. I keep having a...
01 August 2024 7,199 1 View
I am using the Polyomics Integrated Metabolomics Pipeline (PiMP) for metabolite feature analysis. After annotating my data, I have encountered cases where multiple potential metabolites are...
24 July 2024 5,789 0 View
What cell proliferation assay do you suggest? cyQUANT from thermo Fischer has good reviews, a lot of good articles for the kit from Abcam. and Promega kits look easy and straight forward.
21 July 2024 1,554 2 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
Hello, Does anybody have idea about any software or database that can give information about the possible phospho sites located in a protein? I am trying PhosIDP but its not working or loading...
15 July 2024 1,940 3 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