Bhaskar Chatterjee

10 Questions 14 Answers 0 Followers

Questions related from Bhaskar Chatterjee

Has anyone used the Lonza Primary hepatocytes ( plateable) for drug studies?

Has anyone used the Lonza Primary hepatocytes ( plateable) for drug studies? if so, do you strictly follow the Lonza protocols ( seeding density, culturing conditions etc). When you thaw the vial,...

12 November 2024 3,407 0 View

Has anyone done transfections on HLEB3 cell line successfully?

I need to transfect some plasmids in the HLEB3 cell line but I do not find evidence that this was tried, apart from the transfection of SV40 T antigen in primary HLE cells. If anyone has done...

14 August 2021 306 3 View

Failure with Gibson assembly for one fragment assembly (5.7 Kb backbone with inserts varying between 0.9-1.5kb) ?

I am trying to perform a hifi assembly(NEB HIFI master mix) of a backbone (5.7kb) and inserts ( 0.9 - 1.5 kb) in a single fragment reaction assembly ( a different construct corresponding to each...

16 May 2019 9,948 3 View

Why mP values in a FP assay are decreasing?

I initially did an FP assay with receptor (A) and a flourescent labelled ATP analog (B). For the first three times, i tried varying B conc for a fixed A concentration. I found increase in mP...

08 April 2019 3,775 1 View

I am planning to use cycloheximide for inhibiting protein synthesis in HeLa cells. What concentration should I use?

I need to know what concentration of cycloheximide can be used in HeLa Cells without killing the cells. According to Santa Cruz Biotech that sells this compound, IC50 value of CHX is 532nM for...

24 November 2015 1,077 5 View

Which ATPase assay is better: Malachite green assay or LDH/PK coupled assay for my protein of interest?

Hello, I have a protein whose kd value for ATPase activity is pretty low ( 400-500uM). It is a weak ATPase. Which of the two assays would be more effective for measuring this activity. Also, i do...

28 July 2015 3,714 12 View

Does heat shock to Hela cells produce certain chemicals or changes in cell morphology that affect the MTT assay?

After a certain incubation period at high temperature, I am getting higher MTT readings as compared to smaller incubation periods at the same temperature. Cell viability should ideally decrease...

21 May 2015 200 2 View

Can I use opti-mem if it is accidentally kept at -20deg ?

Actually, immediately after the delivery of Opti-mem (Gibco), it was accidentally kept at -20Deg C. It was not opened. So if i use it now for transfection media preparation, would it affect that...

23 April 2015 4,252 3 View

Does anyone have any idea how stable geldanamycin is at high temperature and high pH?

Geldanamycin is a well known Hsp90 Inhibitor (N-terminal). Does anyone have any idea how stable it is at higher temperatures (say 45-50 degrees) and pH above 7. I have to carry out some...

26 November 2013 3,769 1 View

Can the bacterial cells retain the plasmid but loose the gene of interest ?

I always make fresh glycerol stocks from the primary culture and use them for 2-3 months, before subculturing and making another batch and store it at-80. I have been doing this for 4-5 times. All...

11 November 2013 1,274 15 View