how to solve the problem of transmembrane protein smearing in SDS and western blot ?
Hi guys, I have an experiemnt where three groups of fish were used, two of them were treated with two doses of virus, while the thrid is a control group. I want to compare the means of gene...
06 July 2019 3,718 5 View
I am producing a recombinant protein in Rosetta, my protein is fused to MBP+his-tags at the amine terminus. I have solubilized it with sarkosyl. I am using IMAC column to purify my protein and...
02 March 2017 1,285 9 View
is it necessary to remove the sarkosyl- my r-protein is solubilized in sarkosyl- before mice immunization?
02 March 2017 6,722 2 View
I will purify my r-protein (produced in E.coli) and cleave the MBP and His-tag off it and I have heard about the inclination of the r-protein to aggregate/precipitate when the tag cleavage takes...
02 March 2017 5,644 1 View
My r-protein (in e.coli) is only solublized in urea and I want touse it in a downsteam immunization procedure, dentaured protein is not convenient for production of Ab, how to solve this problem ?
02 March 2017 9,202 2 View
I have a question regarding to your Amylose resin kit https://www.neb.com/products/e8021-amylose-resin I read in the protocol...
01 February 2017 9,668 7 View
I am producing my MBP+his-tagged r-protein in Rosetta (75 kDa). I am using the anti his-tag column t purify my protein. I collected the sample flow-through after passing through the columns in...
01 February 2017 3,961 12 View
Hi guys, I want to ask about RNA isolation procedures from fish skin tissue samples? a full protocol would be appreciated.
02 March 2016 5,239 3 View
Is it tricky to use MS to sequence a small recombinant portion of salmon IgT?
31 December 2015 2,236 0 View
I have a question regarding the result of MS (protein sequencing). We made the recombinant r- IgT (unique immunoglobulin in fish) in mammalian HEK cells, we want to characterize the r-protein, we...
11 December 2015 6,059 6 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
I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...
07 August 2024 505 3 View
Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...
06 August 2024 5,373 2 View
I ran a SDS-page of a bacterial lysate and I want to quantify protein concentration in a specific band. I was thinking of using a standards ladder or make some standards are different...
05 August 2024 9,805 3 View
Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC
01 August 2024 967 3 View
I am using CuBr/THPTA for a click reaction in total cell lysates. I am facing issues with my protein sample in non-reducing SDS-PAGE where it's not migrating properly and most of it remains at the...
29 July 2024 950 4 View
I'm having problems with ion exchange chromatography. After applying my sample (a fluorescent protein), a preferential path inevitably forms. The matrix I use is DEAE-Sephacel. Could someone...
27 July 2024 6,605 0 View
I use 0.5% sarkosyl to solubilise recombinant proteins contained in Inclusion bodies of E. coli. sarkosyl very effeciently solubilizes proteins from Inclusion bodies and I am trying to use...
25 July 2024 5,001 2 View
I am performing IgG purification and I have to show my results on SDS-PAGE. I use 10% tris glycine gel and prepare the samples under non reducing conditions. I am new to antibodies and therefore...
23 July 2024 6,664 6 View
"The Correlation between Nutrition and Transport Mechanism under Abiotic Stress in Plants: A Comprehensive Review" by Muhammad Saleem, Jianhua Zhang, Muhammad Qasim, Rashid Iqbal, and Li Song,...
23 July 2024 9,109 0 View