Madhuparna Chakraborty

10 Questions 6 Answers 0 Followers

Questions related from Madhuparna Chakraborty

Can anyone suggest me a yeast nuclear fraction specific antibody apart from H3??

I am isolating yeast nucleus to check the level of my protein of interest in nucleus. I am checking the purity of my nucleus fraction with anti H3 antibody.Though I am getting a intense band of H3...

26 April 2023 7,370 0 View

Why am I getting histone 3 band in my mitochondrial fraction??

I have isolated yeast Mitochondria following a standard protocol. But while checking the purity of mitochondrial fraction by Western blotting, I am getting a histone 3 band in addition to a hsp60...

25 April 2023 2,702 3 View

Why the data that I am getting in my yeast spot dilution assay is not the same with liquid culture growth curve assay??

I am checking toxicity of my desired protein in two different genetic backgrounds in yeast system by Spot dilution assay. The observation which I am getting is not similar to the data I am getting...

29 November 2021 1,398 0 View

How many colonies of monoclonal cell population is sufficient for screening??

I have been trying to make a monoclonal cell population with my desired gene stably integrating in host cell genome with lentivirus system.. I have got five individual colonies by serial dilution...

19 October 2021 881 0 View

Is it necessary to remove the MTT medium before adding the DMSO for accurate measurement in MTT assay??

I am doing MTT assay of TDP-43 transfected HEK293T cells in a 6 well plate. I am dissolving the MTT in DMEM media without Serum and Phenol red. After adding 850ul of the MTT solution to the cells...

05 April 2021 1,025 4 View

Why am I not getting considerate amount of annexin v and pi staining in TDP-43 overexpressed cells??

I am transiently transfecting HEK293T cells with pEGFPC1 WT TDP-43 construct using PEI transfection reagent. After 40hrs of transfection, I am staining the cells with annexin v apc and Propidium...

30 March 2021 5,066 5 View

Why is there cell debris are coming after 2nd or 3rd passage of the cells???

Our lab is facing a difficult problem for almost a year. After thawing the cells are okay, but after 2nd or 3rd passage there are debris present between the cells. We use mycoplasma removed cells...

18 January 2021 5,957 6 View

Can i use overexpressed myc tagged hsp60 as a mitochondrial fraction control in western blot???

I have cloned Yeast hsp60 with a myc tag in a plasmid with a gal inducible promoter in Yeast system. I have overexpressed this myc tagged hsp60 in all my samples. Can I use myc tag specific...

05 October 2020 4,657 0 View

Can I do four point mutations with one primer??

I want to mutate serine 409 and 410 into aspartic acid. For that two bases have to be changed for converting one serine into aspartic acid. So in my case four bases will be changed and between two...

19 August 2020 3,561 4 View

I have been working on Shsy5y cell line for a while.. Recently there is very much cell debris coming in the cells.. Can anyone give me an suggestion?

I haven't changed anything in the protocol for cell culture..

21 May 2019 9,715 6 View