Proteasome inhibitor (MG132) working concentration?

Hi. I am on experiment with proteasome inhibitor ( MG132 ) on A549 lung cancer cells. I treated 10uM of MG132 for 4, 24, 36 and 48hrs. However, it seems not to work. cells were less dead even...

02 March 2018 3,807 2 View

Is it immunoprecipitation band shifting?

Hi I am trying immunoprecipitation by protein G. I tagged antibody( Rabbit) and I western blot using interesting protein antibody(mouse) I got a result lower picture. but it looks a little...

01 February 2017 5,864 1 View

How to purify linear plasmid for transfection after enzyme cut?

hi i want to make stable cell. i use A549. i tried to do transfection on plasmid. but it is difficult to make it. so i tried to make linear plasmid before transfection. but i am not sure i have to...

07 August 2016 4,952 3 View

ELISA; Streptavidin Coating in PCR tubes or 96-well plate?

09 October 2015 6,281 7 View

How do you make 3M KCl?

i am trying to make 3KCl solution for storage buffer of ph meter. but the powder of KCl doesn't solve in water, i tried to stir it on hot water and do autoclave to increase high pressure and...

03 April 2015 9,679 13 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 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

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 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

How to make a time series imputation benchmark ?

Hello ! In a work, I have to benchmark different algorithms to fill in missing values in time series. I insist on the fact that this is imputation and not forecasting. In my case, I have access...

24 February 2021 7,470 7 View

How to interpolate from a time-series database ?

Hi, I am having trouble with a problem, in the field of Optimal Control and the generation of optimal time-series. Let's consider a system, whose dynamics are represented by dx/dt =...

24 February 2021 9,105 2 View