Philip Spear

11 Questions 11 Answers 0 Followers

Questions related from Philip Spear

How can I generate a similarity matrix for protein sequences?

Bioedit can output an identity matrix, but not similarity. http://www.bioinformatics.org/sms2/ident_sim.html generates a list of similarities. Compiling Clustal omega is taking a lot longer than...

10 October 2018 1,829 1 View

Is there a signal peptide sequence to send a protein complex to the outer plasma membrane?

I'm hoping to express several proteins to form a complex and get exocytosed or put onto the outer plasma membrane. Is there a signal peptide or some process I can exploit to get the whole...

05 May 2018 2,722 2 View

Do gold or silica nanoparticles have high levels of DNA transfection?

I'm having trouble finding head-to-head comparisons of lipofectamine vs nanoparticles like silica or gold for transfecting high levels of DNA into cells in culture. What's the best way to get...

01 January 2018 5,015 3 View

What is the cheapest eukaryotic cell to culture?

In terms of cell culture media and other consumables, what's the cheapest eukaryotic cell to culture? FBS is expensive, serum-free alternatives for cells that require FBS are even more expensive....

12 December 2017 8,597 3 View

How can I dissociate focal adhesions but not cell-cell junctions in cultured cells?

I'm trying to grow a confluent layer of epithelial cells on a filter, then would like to disperse the layer off the membrane. I don't want individual cells to disperse, I want the cell-cell...

03 March 2017 8,670 1 View

Can pluripotent cells be grown into flat confluent monolayers?

I'm unfamiliar with ESC culture. Can they be grown into confluent monolayers? Do they form epithelial adherens junctions?I'm having trouble sorting through the literature, as pubmedding those...

12 December 2016 8,607 1 View

What's the best way to make a stable cell line with two inserts?

I'm trying to make a stable cell line expressing two transgenes. I have no experience generating a stable line. If I insert linearized plasmid with some selectable marker, I understand that could...

09 September 2016 936 2 View

Can you culture hacat cells in a monolayer epithelium on matrigel?

I'm hoping to get a single, confluent epithelial layer of hacat cells growing on top of a layer of matrigel. I can find people culturing hacat cells IN matrigel, but not a smooth layer of each.

04 April 2016 1,398 1 View

How can I quantify focal adhesions in mouse embryo cross sections?

I'm trying to quantify focal adhesions in mouse embryo cryosections, E8.5. We have vinculin antibodies, I'm starting to test them. I can't find any published cross-sections of vinculin staining,...

08 August 2015 1,587 5 View

Why aren't my MDCK cells on polyacrylimide getting confluent?

I'm growing MDCK cells on polyacrylimide gels. I am using sulfo-SANPAH to crosslink collagen to the PAAM. Link included to some pictures I took... on my phone through the eyepiece. There are...

11 November 2014 1,720 6 View

How to get 100% DiI and DiO staining?

I'm hoping to stain two cell populations then mix them together. I'm aware that crystals will mosaically label cell populations, I'm hoping to stain all cells to identify which population they are...

01 January 1970 8,687 4 View