Ragan Pitner

5 Questions 17 Answers 0 Followers

Questions related from Ragan Pitner

What's up with my plasmid prep?

Hi all, I recently ran a restriction digest check of several of my plasmids and one of them was weird. All the plasmids in this picture received the same restriction enzymes and are all loaded...

13 July 2021 2,964 17 View

How long can ELISA plates sit in blocking buffer?

This morning (Tuesday) I started blocking some coated ELISA plates but I was unable to acquire sufficient sample to actually do the ELISAs today. They blocked for 2 hours at 37C before I moved...

09 March 2021 6,028 3 View

How long do antibodies stay bound to positively-selected cells in culture?

Hello, For one of my experiments, I enriched cells by staining them with a PE-conjugated antibody and performing a positive anti-PE enrichment to obtain a population that was 64% PE+. After 4...

18 April 2020 2,577 2 View

Can you use cryopreserved isolated mouse dendritic cells in CD4 T cell coculture experiments?

I'm going to do some OT-II/DC co-culture experiments and was wondering whether it would be feasible for me to do a few big CD11c DC enrichments, freeze them, and then use them in my co-culture...

13 February 2020 5,534 3 View

If I have an aqueous solution of ~20-amino acid peptide that contains a single terminal cysteine, will these form disulfide bonds?

I intend to use maleimide chemistry to attach a short peptide with a terminal cysteine to the N-terminal domain of another protein. Do I need to pretreat the short peptide with a reducing agent to...

23 January 2020 318 3 View