Hi, I deal with synthetases which are dimeric. SHould I consider their dimeric mol wt before calculating their molarity or should I go with the usual SDS PAGE mol wt which is monomeric in Kilodaltons
After affinity through amylose and TEV cleavage, the protein is aggregated in GPC so I am not getting any active protein out of this. How can I minimise aggregation.
03 April 2018 1,118 0 View
And am using 1:400 dilution for my protein's antibody. With overnight 4 deg binding. Should I make it 1 hour at RT. I increased tween in PBS from 0.01 to 0.05 % . I got some less background. But...
09 October 2017 1,327 0 View
I am doing it with the sigma GST assay kit. My blank buffer consisting of PBS, GSH and cDNB is also giving an absorbance. So can you just give me a detailed description in case am making a mistake.
07 August 2017 8,973 2 View
I was inducing with 0.8 mM IPTG and kept 16-18 hrs post induction at 25 degree..am trying to reduce the temp and check the expression ; also lowering the salt and increasing imidazole...
06 July 2017 8,924 2 View
Am getting a chaperone at around 63 kD . I have my construct in pETM41. Am purifying with amylose resin and getting a chaperone after affinity. Will HIC after first step clean my protein?
06 July 2017 8,910 5 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
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 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
Hello, I was running a 12% SDS Page electrophoresis on few granulosa cell samples and got this result after the ponceau staining. The total protein lysate seem to aggregate at 70 kDa ladder mark...
21 July 2024 5,128 4 View
I have been running native page for FAM DNA substrate ( fluorescence samples) for protein DNA binding reaction. Binding is there but towards the end of the lane , I am loosing signals...
17 July 2024 6,213 4 View
I have a protein complex in which some dimers easily dissociate. What methods can I use to stabilize these dimers within the complex without altering the overall structure of the complex? Will...
16 July 2024 2,757 2 View
I was using Superose6 10/300 Gl column for purification of my target protein. the sample consists of various oligomeric state and i would like to identify the approximate elution volume where my...
15 July 2024 5,157 2 View
Hi, I hope you are doing well, I am working to remove DNA from protein sample by using silica hydroxode magnetic beads. I used different binding buffers but I din't get my protein band in initial...
02 July 2024 5,502 4 View
I want to check my protein expression but the sample (target protein) on my SDS PAGE shows a little difference as compared to the control. I measured the band density through IMAGE J but the band...
01 July 2024 1,841 3 View