Can anybody who is familiar with JC-10 (Mitochondria membrane potential dye) staining?

09 October 2016 2,865 1 View

I want to try to use HeLa cell and lipofectamine 2000 to do transfection?

I am trying to optimize the transfection by using lipofectamine 2000 for 2000 ng of plasmid DNA/ well with different amounts of lipofectamine 2000 (5ul, 10ul) in the 6-well plate. There is a...

03 April 2016 6,490 6 View

I am interested in autophagosome study in cell line like HEK 293?

plese provide the protocol how we can go through transfection of the cell using the GFP tag etc, positive control of the expression system to check the expression of the channel?

01 February 2016 8,529 4 View

Can someone explain how to calculate the cell capacitance?

I am recording the sodium currents in whole cell patch clamp methods in DRG neurons (20 micrometer) from the Axopatch-200B and getting capacitance 1.24 pF in the dial. Should the capacitance be...

02 March 2014 9,962 9 View

Protocol for olfactory bulbectomized rat model

I want to study the effect of long term treatment (14 days and 28 days) of a compound in olfactory bulbectomized (OBx) rats on locomotor activity (open field), aggressive behavior and sprouting...

03 April 2011 9,771 2 View

Narmalized Current

I am affraod to ask if any one suggest me How we can normalized the sodium current recorded from the whole cell configurations and calculation of tau value. Iam normalizing with positive value of...

02 March 2010 3,343 0 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

Is there any published or unpublished statistics on dropout rate (Dose1 to Dose-2) for Covid-19 vaccination in India, US, UK or any other country?

Are there any published or unpublished statistics or papers on the dropout rate (Dose1 to Dose-2) for Covid-19 vaccination in India, USA, UK, or any other country where vaccination has started at...

28 February 2021 4,237 1 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

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 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