Why should skim-milk be used for blocking buffer in western blots?

I have just performed my first western blot and none of my targeted proteins were detected. The only thing visible on the membrane was the prestained ladder. Someone suggested that my blocking...

21 August 2018 5,787 4 View

How to grid downscale CMIP5 climate data grid to 0.5 degree using Matlab ?

08 July 2018 2,235 2 View

Merging many to 1 in SPSS version 25?

I have 2 datasets one is the adult in the houshold and the other is the child(ren). I'm trying to merge the child data to the adult data however there may be more than 1 child to an adult (many...

10 May 2018 4,946 6 View

Why would LB-low salt medium be prefereable to normal LB medium?

I've come across a protocol that says to use low salt LB broth (10g/L tryptone, 5g/L NaCl, 5g/L yeast extract) for bacterial growth curves. Why would it be preferable to use a low salt medium...

18 February 2018 2,048 2 View

Why is the deletion of bcsQ using lambda red recombination unsuccessful?

I have been unsuccessful in knocking out bcsQ in Pectobacterium carotovorum ssp. brasiliense using a three-step PCR method. Plasmids used: pKD4 and pKD46 bcsQ is MinD homologue part of the...

17 July 2016 1,533 2 View

Has anyone seen this strange FSC-A versus FSC-H pattern?

I recently ran an experiment where I divided a mouse tumor into several wells of a 96-well plate. Each well was stained with a different panel of antibodies. In only one of the wells, I got the...

22 February 2016 4,653 6 View

Does anyone have suggestions about cell density, DIV and surface preparation for hippocampal neuron culturing for micro electrode array?

27 November 2014 4,362 2 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 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

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 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

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View