10 Questions 34 Answers 0 Followers
Questions related from Carl Manner
A technician in my lab group froze precious primary cell samples without using a cryoprotectant. My question: what can I salvage, and how best to do it?
12 December 2016 8,701 3 View
I'm extracting RNA from PBMCs, and do not know what constitutes a normal yield. What is normal for your lab, what method do you use (TRIzol, RNeasy, etc.), and how many input cells do you use?...
12 December 2016 3,022 9 View
Our lab recently did a test run mixed lymphocyte reaction (MLR) using normal human PBMCs. Medium composition was HyClone RPMI-1640 +10%hiFBS +P/S +Fungizone +2.05 mM L-glutamine. The feeder cells...
10 October 2016 700 4 View
I study two protein isoforms of WNT5A that differ from one another by only 18 amino acids post-processing. These isoforms are highly conserved among vertebrates. We see no evidence that these...
03 March 2016 6,674 3 View
I've been blotting samples derived from cells that were treated with a ligand. I expect this ligand to activate a signaling pathway, and I expect certain components of this pathway to be...
02 February 2016 4,341 5 View
I'm getting a ton of non-specific bands in my westerns. My procedure is as follows, and I'm using 25 ug of protein per lane - the bare minimum I think I can get away with. 1. Ponceau S stain 2....
01 January 2016 8,266 7 View
A question from my PI:"We would like to search a RNA-seq database(s) using a sequence to determine what tissues, cells etc a particular transcript is expressed in. I looked on the NCBI and UCSC...
12 December 2015 5,562 3 View
I'm blotting against STAT3 and pSTAT3Y710. My pSTAT3 antibody works great, but I observed a negative band (a white band that is lower intensity than background) at the predicted MW for STAT3 when...
11 November 2015 2,602 7 View
I have some mean amount of DNA from my ChIP assay (A), and some mean amount from the IgG (B). Call fold enrichment C. C=A/B I have standard deviations for both A and B. However, If I calculate the...
06 June 2015 1,602 6 View
I need a protocol for dissociation of human spleen and lymph nodes into viable single cell suspensions. If you have one, and wouldn't mind sharing it, I would appreciate it very much.
01 January 1970 5,934 2 View
