Deepak Deepak

8 Questions 4 Answers 0 Followers

Questions related from Deepak Deepak

What are the black spots in my E.coli Bl21DE3 pellet after protein induction?

Hi all. After growing and inducting the E.coli culture, I extracted the cells by centrifugation, but there are some black spots in the brown pellet. I was wondering if you observe these black...

13 May 2022 9,258 2 View

How to generate topology file for ligand for Gromacs?

Dear all, I converted a PDB file into Mol2 file using Openbabel (H added) and then fixed the bond order number using this command: perl sort_mol2_bonds.pl jz4.mol2 jz4_fix.mol2 When I try to...

23 December 2020 497 4 View

Can I reuse the NAP-5 colums(GE)?

Hi all, I am using NAP-5 disposable columns from GE for removing DTT from my protein. Can I use the same column again for the purpose of removing DTT. Can you please mention the protocol for...

11 December 2020 3,495 5 View

How to reduce (which reducing agent) a disulfide bridge in a protein and how long does it remain reduced?

I want to reduce the disulfide bridge in my 7kDa protein. My protein has only 1 cysteine at the surface and thus makes a homodimer using a disulfide bridge. I want to reduce the disulfide bridge...

25 November 2020 3,369 5 View

Protocol to flash freeze proteins in -80 Deg C?

Hi all, I have a purified protein (57kDa) which tends to precipitate at 4 deg C after a week (conc. 9mg/ml). Can you please share the protocol to flash freeze protein in Liquid nitrogen and...

17 September 2020 1,592 5 View

Adjusting the pH of KHPO4 buffer after addition of imidazole?

Dear all, I am making a 10mM KHPO4 buffer (pH 7) for protein purification. I need to add 350mM Imidazole to it for the purpose of protein elution, but after the addition of Imidazole pH jumps...

24 April 2020 1,194 4 View

How can I align/superpose an organic molecule on to the DNA which is bound to the protein?

Hi all, I have a protein to which a DNA is bound (pdb file). I want to align/superpose an organic molecule on to the top of DNA in that protein. Please let me know which software can do this and...

05 April 2019 2,428 3 View

Protocol for DNA Isolation from mammalian cell lines like Hela, HeK293T, Mcf7. WITHOUT USE OF ANY ISOLATION KIT.

I want to extract the DNA from these above-mentioned cell lines by using the organic solvent method. I don't want to use any isolation kit from any brand.

01 January 1970 1,985 6 View