15 Questions 12 Answers 0 Followers
Questions related from Mark O. Pekarsky
Hello, I am wondering if there's a "limit" for subculturing HeLa cells? If yes, what is it? Hom many times they may be subcultured? If there's a limit - WHY is it like this? What's wrong with...
18 August 2016 8,920 3 View
Hello, I am experiencing problems with expressing GOI2 in GOI1-IRES-GFP:GOI2 cassette in CD4/CD8 cells separated from PBMCs using Miltenyi beads. However expression of the same GOI2 (using the...
27 April 2016 6,770 1 View
Hello dear colleagues, I am experiencing strange problems when performing transDUCTION on my CD4/CD8 cells (separated using Miltenyi beads from total PBMCs from a buffy coat). For now on it is...
19 April 2016 9,444 1 View
Hello everyone, we are trying to transduce T-lymphocytes (TLs) with vector containing IRES, and cannot see any expression using flow cytometry (our target protein is conjugated with GFP). The...
15 March 2016 5,893 3 View
Hello, in my expression cassette I already have 2 proteins that are expressed using IRES. However I'd like to express another protein at the same time (a homing molecule), but, honestly, I do not...
03 February 2016 5,948 3 View
Hi, We would like to inactivate the activity of biotin added to cell culture, but I am wondering what would be the best way to do it... Normally we have a solution of biotin (powder dissolved in...
11 July 2015 6,377 7 View
Hi I am new in the field so I require expert's help :) Briefly the idea of experiment is that the construct (1/2/3) has to be expressed on the surface of the cell, and with the add of component...
25 June 2015 1,736 4 View
Hi all, Sorry I am new to the field, have never performed that assay before. We have T-cells that express a certain receptor on its surface, and we're interested if it can do something to CD19...
19 June 2015 1,413 4 View
Hi, I received a blood bag for PBMC isolation with subsequent T-cells triage required. I received it yesterday evening and put the bag at +4C. I wonder if I can do a ficoll etc NOT the next...
06 May 2015 9,483 8 View
Hello, I am planning to perform a siRNA knockdown using lifetechnologies Lipofectamine RNAiMAX reagent, so taking into account they require 60-80% confluence of cells on the moment of transfection...
02 March 2015 3,516 6 View
Hello, I am trying to do an IP and I am planning to buy some inhibitors (like in the topic, protease/phosphatase/RNAse..). Can you recommend some "good" inhibitors? Thank you!
20 January 2015 1,934 3 View
Hi, I've been using nanodrop for a long time, however I ised it only for RNA/DNA concentration analysis. In fact, there's a "protein" option in nanodrop that I have never used before, though one...
14 January 2015 9,472 19 View
Hello, colleagues. This is my first time I am trying to grow a cell culture from ATCC. So it is Tera-1 (HTB-105) cells and what they recommend on their site is that the medium has to be the McCoy...
18 November 2014 723 3 View
Hello, friends I will need to prepare some total RNA from cells, however I have never did it before. Everyone says that I need to do it in an "RNAse free zone". What that should be in your...
03 November 2014 1,026 3 View
Hi, friends. Would you be so kind to propose the analog of CRL-7800 cell culture (from ATCC)? We badly need these type of cells to have a hiwi expression in it as a positive control (as described...
28 October 2014 6,718 2 View
