Rakesh Kumar

15 Questions 37 Answers 0 Followers

Questions related from Rakesh Kumar

How to store gel for MS analysis?

Please suggest, how to store a SDS-PAGE gel, after colloidal coomassie staining? I have to do LC/MS analysis, but instrument is not available for few days. can i store the gel? if yes how long? in...

06 June 2017 1,507 5 View

How we can analyze electron density map of cryo EM structure?

how to obtain electron density map of a particular cryo EM structure? And how to analyze it? Is it posiible to see a particular amino acid side chain interaction using cryo EM data? If yes, How?...

12 December 2016 2,053 4 View

Why the protein is not denaturing in SDS PAGE?

I am working with a protein of approx 35kDa, in SDS PAGE the band is coming at approx 72 kDA. If it is in Dimeric form then why it is not coming as monomeric form. Laoding buffer contains 5% BME,...

10 October 2016 8,677 8 View

Had any body done Circular Dichroism studies of ATP bound proteins?

I want to study conformational / structural change in the protein after ATP binding ( I am working with a yeast protein that shows ATPase activity) so I want to see and compare the binding effect...

09 September 2016 4,437 4 View

What is the minimum concentration of protein required to get Circular dichroism data ?

I am working with a protein of approx. 45kDa, I want to take spectra in far UV region, protein is highly prone to aggregation so I am getting very low conc. Please suggest what is minimum conc....

07 July 2016 8,397 5 View

Does the ratio between CD spectra at 208 and 222 nM Matters?

For Circular dichroism studies, we know that if the negative elipticity at 208 and 222 are more then it would be alpha helical protein. but if it is more at 208nm or more at 222 nm, what does it...

07 July 2016 2,475 4 View

Can any one tell me why my RT (reverse transcription) is not working?

I have RNA, isolated from Yeast cells (By TRI (SIGMA) reagent), quality is good after Dnase-1 treatment (on Gel), but cDNA is not formed. MMLV RT is working fine for other strains (RNA samples)....

04 April 2016 9,634 9 View

Does the anti-TAP antibody detect CBP tagged protein (Calmodulin binding peptide)in Western Blotting?

I am using a fission yeast strain which has a Calmodulin binding peptide(CBP) tagged protein.I want to check whether this CBP tagged protein interacts with my protein of interest by CoIP. I plan...

10 October 2015 8,765 2 View

Why does the protein not bind to PVDF/ MCE membranes? Did anybody face this problem in filter binding?

I am doing RNA-Protein interaction exp. I want to see binding by filter binding assay, I used Millipore PVDF 96-well filter plates, with vacuum manifold, but could not see binding, I also tried...

08 August 2015 2,484 4 View

What are the importance of these compounds in synthesis of drugs....1-Indanol , Methyl-4 pyridine methanol , 1-Naphthyl)ethanol ?

What are the possible applications of these compounds..1-Indanol , Methyl-4 pyridine methanol , 1-Naphthyl)ethanol?

07 July 2015 9,065 0 View

What could be the reason for protein degradation during sample preparation?

During sample preparation for SDS-PAGE, some of my samples are precipitating metimes. They give smear like band on gel, what could be the reason? Heating at 95 for 5min and then cooling on ice for...

02 February 2015 6,996 7 View

How do I get rid of protein aggregation?

A Splicing factor Protein that is prone to aggregation at/ above 0.5 mg/ml concentration. It also showed temperature dependent aggregation above 65 C. I need to do some Near UV CD studies, as well...

02 February 2015 5,771 5 View

Can anyone tell me? Why the peak shifts in fluorescence measurement of a protein sample?

I have often noticed red shift after moving from high to low temperature.

02 February 2014 6,074 3 View

What could be the solution, if a protein aggregates above 50 degree Celsius during spectroscopic measurement.?

The protein is in 10 mm buffer and concentration is low (25 micro gram/ml)

02 February 2014 9,429 8 View

How to avoid antifoaming agents during fermentation?

For large scale production, I had set the conditions similar to shake flask, but the cell growth had been restricted (evident from abs-600nm- 1.2). What could be the possible reasons?

04 April 2013 2,675 2 View