Ketul Saharan

7 Questions 9 Answers 0 Followers

Questions related from Ketul Saharan

ADP bound in the active site of AAA+ ATPase?

Hello everyone We are building an atomic structure model against a cryo-EM map of AAA+ ATPase. Considering that AAA+ polymerizes when ATP is added, we collected data for AAA+ mixed with ATP. Our...

05 April 2024 4,672 2 View

How to link seperate domains (chains) built for cryo em map?

Hi all, I am working on a cryo em map of a multi domain protein. I am using alpha-fold structure model to fit in the cryo em map. However, due to conformational flexibility the whole multi-domain...

09 December 2023 5,915 1 View

Sample prepration guidlines for cryo-EM of a multimer forming(heterogeneous in form of oligomerization) Hexamer protein?

I am planing for cryo-em of a AAA+ ATPase (natively hexamer of 360 Kda) properties> forms multimer of hexamer at concentration above 2mg/ml and below Nacl 500mM. Also show denaturation on...

16 October 2018 9,810 1 View

Why more bands are coming in pellet(dissolved in 8M urea) fraction rather than soluble fraction after SDS PAGE?

I was doing expression test for a protein. Of gene which I cloned in pETM41 vector and expressing in E. Coli system. When I grown the culture at 20 degree Celsius induced with .5 ,0.25, .125 mM....

16 June 2018 2,502 9 View

Why my protein shown clotting when i added PMSF?

Hello, I was doing limited proteolysis of a protein to purify certain domain and i have optimized with 500:1 of protein to protease (trypsin). i took 2000ug(6mg/ml) (333ul) protein and 4 ug...

10 June 2018 7,534 8 View

Why i am not getting any amplification and band in agarose gel after doing Site directed mutagenesis PCR to generate point mutant?

I have cloned a mycobacterial gene in pETM41 vector, in E coli EXPRESSION SYSTEM, protein expression was there but it is 44 kDa instead of 86 kDa because of premature termination during...

08 June 2018 1,835 4 View

How long i can store bacterial culture in LB media at 4 degree Celsius?

I have bacterial culture of BL21 DE3 transformed with pET28a containing the gene of interest, stored in 4 degree Celsius from 4 months. Do i use culture stored at 4 degree or glycerol stock stored...

27 May 2018 390 6 View