Akhilesh kumar Gupta

6 Questions 6 Answers 0 Followers

Questions related from Akhilesh kumar Gupta

I am getting multimeric band in untreated glutaraldehyde recombinant protein in glutaraldebhyde crosslinking why ?

I was doing glutaraldehyde crosslinking experiment of recombinant protein , i am trying to crosslink at room temerature but I am getting more than one band in untreated glutaraldehyde

08 August 2025 6,386 1 View

Can we used clear bottom Corning 96 well plate for spectrophotometry enzyme assay in far uv wavelength of 260nm ?

I am working on drug target, I have to do spectrophotometry enzyme assay and enzyme kinetics of this protein in 260nm. So I don't have uv transparent plate. So, can we use clear bottom Corning...

22 February 2025 8,685 1 View

During dialysis, my protein get precipitated at 4 degree in cold room codition and why ? anyone suggest me, How can get out from this problem ?

After purification,my protein getting precipitated. for Cell lysis, equilibriation , wash buffer and elution buffer , used 20 mM tris HCL of pH 7.8. I set up dialysis in different percent of...

21 October 2024 4,988 1 View

In sds page and western blot, non specific expressed protein with very thin band is also there along with thick band of specific protein. why ?

Asper discussion, First time my clonning was failed becuase the first 45 nucleotide was not cloned in pet30 a vector then I choosed the different clone of 1:1 ration fraction, I got the size of...

14 June 2024 7,696 7 View

I cloned a gene whose size is 483 base pair, on the basis of sequencing result first 45 nucleotide from start codon is not cloned in vector why ?

NdeI restriction site was added in forward primer and HindIII was added in reverse primer size of band is looking near 500 bp but size of is less than 15 kda but expected size should be 17.7kda.

16 April 2024 269 5 View

I transformed a gene in E.coli DH 5 alpha cells after ligation in pet30a vector, as a result I didn't saw any separate clone they all are merge

please anyone tell me why this happens

01 January 1970 6,305 2 View