Have I to consider the oligomeric status of my protein before calculating its molarity?

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

11 December 2018 3,287 0 View

Hello, my protein is undergoing aggregation and eluting in void volume during gel filtration. How can I reduce this ?

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 1,046 0 View

Hello < I am trying to standardize IFA with Plasmodium falciparum parasitized RBCs . Can you help in that?

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 1,251 0 View

Can you give me a protocol for GST assay?

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 8,894 2 View

Does HIC help to get rid of chaperones?

06 July 2017 8,799 5 View

No title

02 March 2021 3,060 3 View

12% ledeburitic cold work tool steel, etched for revealing retained austenite, any help for clarifying?

Hello everyone. Given microstructure image belongs to a 12% ledeburitic cold work tool steel (annealed and tempered). The sample was etched with picral (100 mL ethanol and 4 g picric acid). I...

02 March 2021 4,119 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View