Why can't i get ladder resolution?

I attach a photo of my agarose gel.. As you can see the DNA ladder runs like "all toghether" (sorry for bad english). I followed the manufacturer recomendations (1,5% agarose, 90mV, 1 hour)... The...

05 June 2016 4,582 11 View

Why didn't PCR work?

I'm doing RT-PCR from rat catilage tissue, and the thing is i searched for tissue specific markers such as SOX9, Aggrecan and type 2 collagen. this is what it looks like, the last 3 lanes are the...

03 April 2016 7,328 10 View

Why do i get a smear on PCR???

01 January 1970 2,452 0 View

How can i get rid of bright background on agarose gel??

I'm running my pcr products in an 1% agarose gel made with TAE buffer. The volume i use is 20 ml with 1ul EtBr. I attach a picture. I want to get as dark as posible for the background and as...

01 January 1970 9,009 11 View

BSA-AGE without centricon?

i need to prepare BSA-AGE for my stem cells culture. The problem is i have centricones 100 kDa, i the ones i need are the 10 kDa ones. Is there another protocol for preparing BSA-AGE without...

01 January 1970 9,775 3 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

No title

02 March 2021 3,060 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

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 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