Can protein A magnetic beads be used for depletion of cells?

I am aware of Streptavidin magnetic beads which can be used with biotinylated antibody in MACS. But I would like to know if anyone has tried isolating or depleting cells from heterogeneous...

05 June 2016 8,652 2 View

How to cryopreserve mESCs expanded in 2i medium?

02 March 2016 4,026 5 View

Should the normalisation of microarray data be done using all samples or on pairwise basis (ctrl & test) for differential expression analysis?

I would like to know if it will be correct to perform normalisation of raw microarray data on every pair of samples to be compared (eg, control vs treatment 1, control vs treatment 2, etc) and...

10 November 2013 554 6 View

Detection of APC protein?

I have been trying to detect APC (adenomatous polyposis coli) from RIPA lysates of HEK and SaOS2 cells by western. However, I am not able to detect the protein (Mol Wt ~300kDa). I have tried...

02 March 2013 8,144 8 View

Cells detaching in culture?

I am facing a problem with all cell lines and primary cells I am using like hMSCs, SaOS2, HEK, NIH3T3. Since past about a week, the cells have started to detach from the surface and this...

04 May 2012 1,926 92 View

MSC transduction

Hello everyone, I am trying to optimise MOI for transducing hMSCs using TurboGFP lentiviral particles (from Sigma). I incubated cells for 16h at MOI 1,5 & 10, and analysed after 48h by flow...

05 June 2010 5,272 2 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