Furin western blot - 2 bands?

I am expressing furin gene in a furin deficient cell line (LoVo), but I always get 2 bands on my western blot ~ 87kDa, corresponding to furin expression and a lower band, of approximately 50kDa,...

08 September 2018 1,754 0 View

MRNA maturation after transfection?

I am having some problems trying to clone a mRNA, maybe due to it's amount compared to other mRNA. I have tried PCR using cDNA from oligo DT, specific or random primers, but with no success. I was...

07 August 2018 3,497 3 View

Contamination after hygromycin supplementation?

I am trying to establish a stable cell line after transfection with pcoHygro plasmid. Every time I supplement with the selection antibiotic (hygromycin) I get some small contamination (it seems...

06 July 2018 9,790 6 View

PCR using oligo dT?

Is it possible to perform a PCR, using as the reverse primer a oligo dT, the same used in the reverse trancription? I am trying to amplify a mRNA, but with no success

05 June 2018 6,831 4 View

Mouse vs rat?

I need to produce antibodies against my protein in 2 different animals, amd my university provide us mice and rats, but I am not sure if the secondary antibody will cross react between those...

01 February 2018 784 3 View

Baculovirus protein exporting pathway?

I am expressing a protein using Baculovirus / SF9 cells system, and I was wondering if, with the signal peptide for exporting the protein (pAcGP67a plasmid), the protein will go through Golgi...

09 October 2017 6,361 0 View

I am working with MinION sequencing technology, with a RNA imput. It generates several read that can not be assembled. Any clue how to analyze them?

The reads are not assembled as contigs, and I have tried some different softwares with no success, such as EPI2ME and Geneious. I have no idea how to align and check my sequences, I have tried...

08 September 2017 2,970 3 View

What is the difference between Vero and Vero E6 cell line?

I've heard that some viruses can only infect E6, and I have no idea why. Anyone

01 February 2016 1,014 0 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

What are the values needed for gabor filter's parameter for cifar10 data?

Hi, I implemented a code to gabor filter cifar10 data but the images after being filtered and stacked are not clear like the original images. I think the problem is in the way I am using the...

03 March 2021 6,317 1 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

How to calculate Total Flavonoid?

I analyzed flavonoid total spectrophotometry, the procedure as below: 2 ml supernatan added 5 ml dd.H2O, then added 0,15 ml NaNO2 5%, 5 minutes later added 0,15 ml AlCl3 10%, After 6 minutes,...

01 March 2021 6,722 1 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 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