Accutase is gentler on the cell surface proteins for detachment so you don't potentially lose the signal.
Someone taught me that when I first did flow cytometry but I've seen people using trypsin anyway.
Just thought I'd share.
Feel free to share other nuggets of knowledge.
My textbook says it is highly specific for R(n-1) = Arg, Lys; R(n) ≠ Pro but mentions nothing about the stereospecificity. I also couldn't find anything online about it. Does it mean is not stereospecific? I think it is not stereospecific, because it's a digestive enzyme, that doesn't need to be that specific to do it's job, but I am not sure about it.
Just diagnosed as having this disease. I inherited one mutated gene on chromosome 14. Been experiencing life long health issues in lungs, liver, skin. digestive tract. Looking for info to help me deal with this disease. Anyone out there who has the genetic disease. Thankfully I have a great team of doctors now. But have been misdiagnosed and delayed diagnosed for decades.
This might be a silly question but am I supposed to see cell debris after stopping my digestion and spinning it down? My protocol says transfer supernatant into separate tube (which I know is where the digested protein is) but I didn’t see any pellet or cell debris at the bottom. Was I supposed to? My samples are being prepared for MS analysis so after lysis I performed a methanol/chloroform precipitation and then subjected my samples (10 ug protein) to trypsin digestion for 20 hours over night. I stopped my digestion by adding 96% formic acid to 5%.
If you know you know.
Hey,
I'm interested in genetics, but currently, I'm just a student. I have a question: I would like to stain chromosomes (G-bands) by treating them with trypsin and staining with Giemsa. Unfortunately, I couldn't find any protocols online to know the concentration of the trypsin or the application medium. Can anyone here help me? I just need like a procedure for the method, including working concentration specifications.
Thanks!
I am working with an adherent cell line and am seeing a lot of cell clumps after fixing, staining, and then filtering samples. Can I add trypsin to the cells right after washing out antibody to declump a little and then wash it out before running them on the flow cytometer? I know that it can be an issue for some surface markers, but thats not an issue for me.
The 14‐amino acid cyclic peptide SFTI‐1 is becoming increasingly recognised as a valuable prototypic peptide and has planted seeds for thought in a growing number of fields. This Review surveys the applications of SFTI‐1 in the design of protease inhibitors that have blossomed since its discovery, and the new shoots that have recently sprouted in the areas of pharmaceutical design, synthetic chemistry, and molecular biology.
Abstract
Nature‐derived cyclic peptides have proven to be a vast source of inspiration for advancing modern pharmaceutical design and synthetic chemistry. The focus of this Review is sunflower trypsin inhibitor‐1 (SFTI‐1), one of the smallest disulfide‐bridged cyclic peptides found in nature. SFTI‐1 has an unusual biosynthetic pathway that begins with a dual‐purpose albumin precursor and ends with the production of a high‐affinity serine protease inhibitor that rivals other inhibitors much larger in size. Investigations on the molecular basis for SFTI‐1′s rigid structure and adaptable function have planted seeds for thought that have now blossomed in several different fields. Here we survey these applications to highlight the growing potential of SFTI‐1 as a versatile template for engineering inhibitors, a prototypic peptide for studying inhibitory mechanisms, a stable scaffold for grafting bioactive peptides, and a model peptide for evaluating peptidomimetic motifs and platform technologies.
https://ift.tt/38xzvL9
I've been having funky results and I've come to the conclusion that the problem is my lysates and not the antibody. In an effort to optimize everything, I've been reading about cell scraping being preferred to trypsin for phosphorylation studies and I'm not too worried about viability as the cells have to be used anyway. Feedback or suggestions would be great.
Hi there,
Wondering if anyone knows of the stability of lyophilized enzyme at RT, specifically trypsin. I’ve tried looking into it before and have had little success.
I left out some trypsin overnight (normally kept in freezer) and am wondering if it will still be fine.
