Pratibha Srivastava

8 Questions 65 Answers 0 Followers

Questions related from Pratibha Srivastava

JMP Pro 14 and one way ANOVA?

I have to analyse the data for one way ANOVA using JMP pro 14 software. Anyone dealt with this software or has any kind of experience. Kindly share it. Thanking you in advance.

24 September 2020 783 6 View

What is the minimum nuclear lysate fraction concentration is required to set up a Co-IP?

My protein size is 240kDa and I am interested in identifying the interacting target protein at endogenous level. I have earlier tried using the whole cell lysate but 4-5 mg of MEF cell lysate do...

22 May 2019 7,194 0 View

Why is my recombinant construct failed to restore the same gene expression level as of endogenous protein?

I synthesised pAdenovirus expressing Protein-FLAG tagged and reconstituted in knockdown cells. When I perform the experiment, WT cells can perform the gene expression but reconstituted cells can...

17 May 2019 3,381 3 View

Co-Immunoprecipitation: Why do I find similar bands in pre-immune rather than interacting region?

I am finding which domain of my protein A interacts with protein B. I have designed a 250kDa protein A into three fragment tagged with HA at both of their terminal. I have tried to pull down the...

20 February 2019 846 1 View

Can anyone explain me how to design tet-on and tet-off system in mouse and human cell line?

Is there a different methodology in mouse and human expression. I need to check the expression of a gene under the influence of two proteins. Thanking you.

25 January 2019 8,277 1 View

Why do DH5 alpha express different length of transformants from a same ligated product?

I experienced a slightly different transformants length of my ligated product. They maybe varied by less than 100bp in length!

13 December 2018 5,945 2 View

Why do the mutants aren't expressed in the 293 cells whereas Full-length WT construct is expressed ?

I prepared the recombinant protein X-FLAG tagged and two of its mutant. I have checked the mutant was correctly assembled in the pShuttle vector but cannot discover its expression even after 1...

11 December 2018 422 16 View

I want to prepare an overexpression module for my protein with a N-ter HA tag. Can you suggest me the possible strategies?

I have tried by breaking the protein into multiple fragment and using the PCR products to clone one by one. I successfully cloned into the pShuttle vector but I couldnt get the expressed protein,...

27 February 2018 9,981 2 View