20 Questions 18 Answers 0 Followers
Questions related from Mark O. Pekarsky
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...
04 April 2016 6,880 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...
03 March 2016 6,599 4 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...
02 February 2016 6,018 3 View
Hello, We are trying to find some biotin conjugates to perform flow cytometry analysis. Are there any nice conjugates used by "people around the world"? Thanks a lot for suggestions! P.S. Found...
02 February 2016 8,287 5 View
Hi, I am trying to perform an expansion of T-cells, purified (by miltenyi columns) from PBMCs and transduced with construct of interest. I am wondering if there's a reliable and "working" protocol...
12 December 2015 425 5 View
Hello, everyone. We all know how, sometimes, it's difficult to write down everything into a lab journal, while I believe there are other approaches apart MS Word :) Would anyone be able to share...
07 July 2015 5,393 0 View
Hello, I would like to know how different are Kds between avidin+streptavidin binding protein and streptavidin+streptavidin binding protein? Thank you
07 July 2015 5,990 0 View
Hi, I am looking for a cell line that can be a negative control for a Raji cell line. Did anyone already tried it? The idea is to use Raji for degranulation assay (to see if T-cells, interacting...
06 June 2015 9,812 3 View
Hi, colleagues. Unfortunately I have just started to work in this new field for me, PBMC purification from blood bags, MACS separation etc. So what I do is out of a blood bag from hospital I do a...
06 June 2015 5,875 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...
03 March 2015 1,460 5 View
Hello, I am about to complete my FACS training so I need to bring some cells labeled with different markers (correct me if I'm wrong in terms). What I have here in culture are Jurkat cells, so I...
02 February 2015 7,404 3 View
Hello, I am about to IP my protein of interest and maybe (I hope so) some small RNA would co-IP with it. Since this is my first time in this area, would anyone be so kind to suggest the best...
01 January 2015 532 4 View
Hello colleagues, This is one of my first IPs so I am a bit confused with the result I just obtained. I used the dynabeads protein G for my purposes. Here it is, my protein is ~97kDa (in the red...
01 January 2015 3,571 18 View
Hi, I expect that my protein binds some RNA that I would like to isolate, so I have tried to IP my protein but it seems that it's not working since commercially available Abs do not IP my protein...
01 January 2015 8,987 15 View
Hello, I am trying to perform some IPs using Dynabeads G from invitrogen on Ago2 protein from Jurkat cells, and I need a negative control. I have some rabbit Abs for my target protein (Ago2), and...
12 December 2014 9,993 3 View
Hello, I am planning to IP and Ago2 protein from Jurkat cells, but unfortunately I haven't been doing an IP before. I am planning to use dynabeads protein G for my purposes and I have browsed...
12 December 2014 4,779 3 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...
11 November 2014 9,746 4 View
Hi friends, I am using the SuperScriptFirst-Strand Synthesis System for RT-PCR, from Invitrogen (#11904-018) and, unfortunately in our kit the RNAse H vial is lost. Of course I can just buy a new...
11 November 2014 8,133 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...
10 October 2014 877 2 View
Hi, Apologize in advance for the incompetence - I am new to that system. I am also new to the field and to the cell work - that project is different from what I have been doing before. I am...
10 October 2014 4,423 3 View
