How to do co culture of human fibroblast and breast cancer cells ?

Hi all, I want to perform the co culture of human dermal fibroblast and breast cancer cells. I have collected conditioned media from cancer cells and treated on fibroblasts and observed the...

10 November 2017 3,628 1 View

Why fibroblasts Change morphology after one/two passages ?

Hello, I am working on human dermal fibroblasts. After thawing from liquid nitrogen, there is clear morphology till one/two passages. But after that we observe a change in morphology, like the...

02 March 2017 1,131 6 View

Can anyone please suggest a normal breast-associated fibroblasts (NAF) and Breast carcinoma associated fibroblast(CAF) cell line..??

Hello friends, Can anyone please suggest a normal breast-associated fibroblasts (NAF) and Breast carcinoma associated fibroblast(CAF) cell line..?? One more query, I have HaCat cell line, can it...

07 August 2016 6,985 1 View

How much PBS should I dilute for preserving exosomes. ?

07 August 2016 8,322 5 View

How to quantify Exosomes ?

I have isolated exosomes from conditioned cell culture media . I have to go for its downstream applications and need to do the MTT assay. For that how to quantify the exosomes. Can we go with...

02 March 2016 4,466 12 View

Can we use incomplete media for exosome isolation ??

Instead of using exosome free FBS in complete media, if we go for incomplete media (after culturing cells in complete media and then washing with incomplete media and flooding the culture vessel...

02 March 2016 936 2 View

Which is the better option for Exosome Isolation ..??

Which one is better to go for Exosome isolation from cell culture media in terms of it's yield ? Exoquick from system biosciences or Total exosome isolation reagent from ThermoFisher sc....?? Yes,...

01 February 2016 6,752 3 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Alginate and silk fibroin hydrogels toxic to cells?

Hello, I asm currently working with alginate and silk fibroin hydrogels. I prepared a blend of alginate and aqueous silk fibroin (straight from dialysis) and resorted to ionic crosslinking to...

23 February 2021 8,249 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Why the nucleic acid gel melted after running for 30min?

Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min. My gel concentration is 1% percentage...

22 February 2021 9,896 3 View

Sanger Sequencing results not matching experimental results of Restriction Digestion, colony PCR and protein expression?

I have cloned two domains of a gene into two bacterial vectors using cut and paste cloning with different RE sites. I checked the putative clones by colony PCR and Restriction Digestion (fallouts...

18 February 2021 6,736 4 View

How to dissolve lyophilized silk fibroin?

Dear members of the community, I am trying to concentrate silk fibroin to 25 % (w/v). Nevertheless, i am struggling with the concentration step. I successfully achieve concentrations of 5-6 % of...

16 February 2021 1,808 1 View