Aniket Limaye

9 Questions 13 Answers 0 Followers

Questions related from Aniket Limaye

Can ethyl acetate extraction of buffered enzyme reaction prior to chiral column HPLC remove Tris salt & NADPH from the reaction sample selectively?

Hi! Using Chiral HPLC column (OB-H), I am trying to analyze final enzymatic reaction products of NADPH dependent aldo ketoreductase for possible formation of asymmetrical alcohols of...

21 August 2021 7,895 9 View

D/L or R/S chemicals, are these racemic? If I purchase such chemicals, can I obtain separate peaks for each orientation of these isomers?

Dear all, I am very new to chiral HPLC. Can someone help me understand a thing regarding chiral HPLC : I am studying stereoselective reduction of bulky ketones by recombinant aldo-keto...

16 April 2021 8,837 3 View

How to find out co factor specificity of aldo keto reductase enzymes?

I have expressed 3 recombinant Aldo keto reductase genes from Gram negative bacterium belonging to the genus Chryseobacterium. Will I have to purchase and try all the cofactors like NADP, NADH,...

24 November 2020 2,858 1 View

If I store β-NADPH stock solution prepared in Na-bicarbonate buffer, pH9 as per SIGMA, can I use this in reaction with pH other than 9 ?

I am planning to make a single time stock solution of β-NADPH (10mM, 100x concentration) from 100 mg desiccated powder and store its single-use aliquot vials at -20C refrigerator. I am a bit...

03 December 2019 5,127 0 View

Is there any way to determine presence of disulfide bond in the protein structure by using only amino acid sequence?

I have to choose an expression host with encoded rare codons for expressing pET vector insert. There are multiple choices of E.coli derived strains available from Novagen. The family of Origami...

08 March 2019 6,585 5 View

Could anyone comment upon growth medium for long term preservation of 'untransformed' E.coli BL21(DE3) and E.coli BL21(DE3)pLysS competent cells?

I am having last few single use aliquotes of untransformed E.coli BL21(DE3) and E.coli BL21(DE3)pLysS competent cells from a used up cloning kit. Is there any specific growth medium or additional...

21 February 2019 4,990 3 View

Are there any cloning limitations for pET32 plasmid if the target protein is about 100-105 KDa ?

I have inserted laccase gene in pET32 vector, verified the sequence, cloning site and transformed it into E.coli BL21(DE3) as well as E coli BL21(DE3) pLysS. I have also tried using carbenicillin...

19 November 2018 1,657 0 View

Why carcinoma cell lines (e.g. HepG2, Caco-2, RAW) are used in invitro cytotoxicity experiments?

HepG2 is considered as a model system for various cytotoxicity experiments according to many studies. Won't it produce wrong results if carcinoma cells have been used for such studies as such...

22 August 2017 5,525 7 View

Is there any standard criterion to select optimum number of sequence candidates for establishing a phylogeny tree in bacteria using MEGA?

for example, in case of 16s rRNA nBLAST results, I have almost 80 similar sequences with % similarity ranging from 99% and going down till 94% ?

17 March 2017 4,951 2 View