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Questions related from Carl Manner
Wondering what the minimum time I can treat my cells for is. Virus is VSV-G pseudotyped. Also, how long until the provirus is fully integrated? Treating sea urchin eggs, and to reduce mosaicism...
16 June 2021 8,738 1 View
I feel pretty silly asking this, but am stumped. I'm attempting to do dCas9-CAPTURE (paper: 10.1016/j.cell.2017.08.003) and am having issues during the chromatin isolation step - the chromatin...
10 March 2020 7,613 0 View
There are tons of protocols for hepatocyte isolation from whole livers and lobectomies, and they all depend on perfusion with a collagenase solution. My group is collecting wedge biopsies from the...
22 April 2017 1,991 1 View
I've isolated human splenocytes. The tissue was dissociated, cells were spun down, red blood cells were lysed, and cells were washed a few times and filtered through 70um mesh. During processing,...
15 March 2017 4,540 3 View
I'm doing a Mixed Lymphocyte Reaction and sorting out alloreactive T cells by FACS. Briefly, I treat feeder cells with 25ug mitomycin C for 30 minutes and wash them 2x with RPMI+10% FBS to render...
23 December 2016 313 7 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...
28 October 2016 8,823 3 View
I'm looking for a marker specific to T helper 3 cells (Th3) for use in a multicolor flow cytometry panel. Any suggestions for a specific marker, or combination of markers, is greatly appreciated.
18 September 2016 3,303 2 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...
28 March 2016 538 2 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...
22 February 2016 1,540 4 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...
14 December 2015 2,201 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...
02 November 2015 6,370 6 View
I have two stable cell lines (in mouse L cells), each of which overexpresses one of two isoforms of WNT5A. From these cell lines, I've generated conditioned medium. Before treating cells with...
23 October 2015 6,934 4 View
So, here's the data I have: Microarrays showing DEGs from cell populations treated with 5-aza and untreated cell lines. Bisulfite seq data for the same cell lines with the same treatments. My...
17 April 2015 3,513 9 View
Wnt5A is a non-canonical wnt ligand that activates one or more of the known non-canonical wnt signaling pathways. One of these pathways is the Wnt/PKC pathway. I need to quantify PKC activation...
03 March 2015 3,193 5 View
