What is the best website or software by which I could determine transcription factor(s) that potentially bind to upstream my gene of interest?

I am trying to find a transcription factor(s) that potentially binds to a specific segment of my gene of interest. The segment is a little bit large (about 3kbp). knowing which transcription...

09 October 2019 2,957 2 View

What does it mean when you have less cells# in flow cytometry plot compared to a control group?

Hi! I have run flow cytometry for 3 different cell lines (black, blue, and red) which were stained with red dye (FL2) with a same event number (15,000 events). However, the cell number (count; Y...

09 October 2017 2,961 3 View

Is there any ideas why the over expressed protein behaves differently than the endogenous protein?

09 October 2016 916 9 View

What is the best way to test several genes expressions using qPCR?

06 July 2016 5,179 3 View

What is a good or official program to quantify western blotting results?

What is a good or official program to quantify western blotting results? Any help would be appreciated and thanks

08 September 2015 1,600 2 View

Does someone know a web sit(s) by which we can check the biological conferences in the USA?

03 April 2015 909 0 View

Is there any way to avoid cleavage of eukaryote protein expressed in E coli?

Hello, I am trying to express eukaryote protein (36 KDa) in E coli ( BL21 and Rosetta) by using different vectors (pGEX-4T-2 and pET-28a) and tags (GST and His- tag, respectively). However, more...

03 April 2015 7,830 8 View

How many times can we use GE Healthcare GSTrap FF, 1 ml?

31 December 2014 4,600 0 View

Is there any way to purified a protein that does not bind in either cation column nor anion column?

Hello, I express my protein with GST tag and trying to get my protein pure without the tag. Therefore, after I got GST-protein partially pure by performing affinity chromatography, I cut the tag...

31 December 2014 4,945 21 View

Which are the amino-sequence cutting sites of bovine thrombin?

Hello, I am trying to cleavage my protein from GST-tag by using Thrombin with its site ( L-V-P-R-G-S) which is provided by plasmid. However, I am confused because I saw after running my protein...

10 November 2014 6,200 10 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

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

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

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why the cells die after stable transfection?

I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...

22 February 2021 2,864 4 View

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 3 View

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

Which transfection reagent is the best for plasmid transfection of MDA-MB-231 cell line, Lipofectamine based or other reagent suggestion?

I try to transfect MDA MB 231 cell line with a plasmid by PEI, unfortunately, the rate of transfection is low, and now I wanna find and order the best transfection reagent for this cell line and...

19 February 2021 828 6 View