How do you use PKH dyes for intra-cellular endosome labeling?

Hi, I want to label adherent brain endothelial cells with pKH26 to visualize endosomes inside it. I tried staining it. However, my pKH26 staining is not consistent as well as it is not...

31 December 2017 3,568 1 View

Have anybody used far red fluorescence dye for staining bacteria for live microscopic imaging?

I am searching for a fluorescence far red dye to stain bacteria for live Microscopic Imaging. PSVUE is one of NIR (Near Infrared Dye) which stains anionic lipids but it is not compatible with PBS...

09 October 2016 1,850 0 View

Good Ubiquitin ligase inhibitor to block ubiquitination in HBMECs In Vitro

HI. I am trying to inhibit ubiquitination in Human Brain Endothelial Cells In Vitro using PYR-41 ubiquitin ligase inhibitor. But it is not inhibiting ubiquitination as lower concentration and at...

07 August 2016 8,242 2 View

IS there a staining for adherent and tight junction protein in fixed cells?

04 May 2015 8,288 9 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

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 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

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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View