Does anyone have experience of transfecting siRNA in hMSC?
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 8,740 2 View
I am growing mESCs in 2i medium. I have cryopreserved mESCs cultured in DMEM+FBS in 90%FBS+10% DMSO but would like to know how to cryopreserve cells grown in 2i medium which is free of serum. I...
02 March 2016 4,117 5 View
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 640 6 View
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 8,255 8 View
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 2,035 92 View
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 5,398 2 View
I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...
03 August 2024 3,851 1 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...
30 July 2024 6,648 1 View
Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....
29 July 2024 5,007 0 View
I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?
29 July 2024 3,229 1 View
I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...
28 July 2024 3,178 3 View
I have a Polyjet that has passed its labeled expiration date for 1 year and I'm going to re-start transfection experiments. It is really needed to change it? Transfection efficiency may be lower?
23 July 2024 8,059 1 View
Hello! We are using LNP to deliver circular mRNA into T cells. However, compared with linear mRNA, the transfection efficiency and protein expression duration are similar. Are LNP generation or...
21 July 2024 4,322 1 View
Im trying to transfect A549 cells using lipfectamine 3000. im using a 96 well plate with 1microgram plasmid DNA in each well . should i remove the culture media or change it or decrease it befor...
21 July 2024 7,610 2 View
Hi All I am looking to get your opinions, I have perfomed liposome mediated DNA transfer into bacteria, the DNA is a plasmid with msfGFP and when I induce the plasmid I see the fluorescence, and...
16 July 2024 4,698 4 View
