Can a single aminoacid substitution renders protein insoluble?

Mutated a Arginine residue with Alanine by SDM in the active site and the protein became insoluble. WT protein was found soluble but the mutated one was found in pellet fraction.

06 July 2016 3,066 4 View

Does Escherichia coli DH5 alpha strain contain fertility plasmids? What is the parent strain?

what is the parent strain for Escherichia Coli DH5alpha and does it have F plasmid?

31 December 2015 5,805 4 View

Do we need to add 0.1% SDS in Transfer buffer for transferring a 97kDa protein?

I did western blotting for a 55kDa and 97kDa protein and used 1:5000 dilution of Primary antibodies and 1:10000 dilution of secondary antibodies. The blot was developed using DAB and i got a good...

03 April 2015 6,737 7 View

Heat exchanger with liquid Nitrogen as coolant?

I want to cool Natural Gas from 195 Kelvin to 138 Klevin in a heat exchanger by using liquid nitrogen as the coolant medium. When entering the heat exchanger, the liquid nitrogen will have a...

01 March 2021 6,450 5 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Alginate and silk fibroin hydrogels toxic to cells?

Hello, I asm currently working with alginate and silk fibroin hydrogels. I prepared a blend of alginate and aqueous silk fibroin (straight from dialysis) and resorted to ionic crosslinking to...

23 February 2021 8,249 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Why the nucleic acid gel melted after running for 30min?

Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min. My gel concentration is 1% percentage...

22 February 2021 9,896 3 View

How to get/count refractive index in gasses?

20 February 2021 8,202 6 View