Is my signal sequence is correct or functional?

My cloned gene contain a signal sequence at N terminal . How can I check the nucleotide sequence is a correct signal sequence. The following uppercase letter are signal sequence cloned in...

01 February 2019 7,597 2 View

Could any one please provide me a mammalian expression vector with AviTag to add at C and N terminal of protein ?

I want to add AviTag with my protein at N and C terminal and to express it in mammalian cell line.

09 October 2018 6,660 0 View

Can I get single fragment using two PCR product in single reaction ?

I am planing to get two gene together for cloning purpose. My plan is first amplify the gene "x" using a set of primer which will amplify gene x without stop codon. I will amplify another gene "y"...

08 September 2018 6,654 1 View

What will be size of my expressed protein If my construct contain Mut. Htt96Q linked with BiRA* ?

My construct contains Htt 96Q followed by BirA* with a 6 amino acid linker between them. Than what will be size of my expressed protein if molecular mass of BirA* is 35.2 KDa? Based on amino...

07 August 2018 3,656 1 View

How long do I need to treat transfected HEK 293 cells with Brefeldin A and the concentration need to see endo H resistance in glycosylated protein ?

I am using knockout HEK 293 cell.

06 July 2018 4,440 2 View

Why I found colony for negative control (contain only double digested vector : one blunt and one sticky end)after transformation in DH5-alfa cells ?

Why I found colony for negative control on selection plate (contain only double digested vector : one blunt (by EcoRV) and one sticky (by SalI) end with ligation mixture, no insert, after...

06 July 2018 231 1 View

I want to ligate my 1kb insert (PCR product) with 6 kb vector, both are digested by ECORV (creating blunt end) and Sal1 (creating sticky end). How?

I want to ligate my 1kb insert (PCR product) with 6 kb vector, both are digested by ECORV (creating blunt end) and Sal1 (creating sticky end). How? Is blunt end may create problem in cloning?

05 June 2018 6,728 3 View

Why Polyst. Flat bot. high Bind. MP (Costar) blank micro-plate is reactive with positive control?

I am using Polyst. Flat bot. high Bind. MP blank (Costar) micro-plate for making ELISA (HIV ), when i am using this blank plate for ELISA assay , it is found reactive with positive control while...

31 December 2016 5,725 6 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Isolates of Hymenoscyphus fraxineus?

Dear Dr. Cai, my name is Simone Prospero and I work in the team of Phytopathology at the Swiss Federal Institute for Forest Snow and Landscape Research (WSL;...

01 March 2021 1,133 1 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View