16 Questions 13 Answers 0 Followers
Questions related from Delilah MH
Hi all, So I want to see if my protein is secreted into the media of my cultured cells and wondering if the correct protocol would be to just collect the media and adding sample buffer to it and...
04 April 2019 5,724 3 View
Hi everyone, I used to see my protein of interest on my blot for all my cell lines but suddenly when I rerun the experiment I don't see it anymore in one cell line! even though I still use the...
03 March 2019 8,184 5 View
Hi, I'm trying to stain my cells after transient transfecting them but each time I get the cells either over confluent, or clumped in the middle even though I swirl the dish in an 8 shape to...
03 March 2019 7,054 2 View
Hi , I have to transfect cells with a GFP tagged plasmid and want to look under the microscope to see the localization of the protein in both transfected and non-transfected cells. How can I do...
02 February 2019 1,039 3 View
Hi everyone, I am newbie on using the confocal and I am wondering if the setting on all the images I take should be the same? in order to compare between them? because when I took the images I...
02 February 2019 7,561 3 View
Hi all, I am doing a transient transfection to my cells with GFP-tagged plasmid and sending them to be FACS sorted and I need to replate them once I get them back. This is my first time ever doing...
02 February 2019 7,496 3 View
hi everyone, I have ben treating cells with a drug for 4 months now and then immunoblotted with AKT as it has been suggested that that drug effects the PI3k pathway. however, the phosphorylation...
07 July 2017 5,107 2 View
Hello everyone, so I have blocked my membrane in 5% BSA then incubated over night with primary antibody in TBST then secondary in 3% milk. when developing I saw the band I'm looking for....
06 June 2017 3,896 5 View
Hello everyone, I have done DNA sequencing on mouse samples after treatment with drug.. i wanted to see if there was any mutations caused by that drug.. I have found some mutations in P53 but...
06 June 2017 10,011 11 View
Hi everyone, I'm planning to sequence my PCR product but first i want to make sure if i prepared my sample to be amplified by PCR correctly. I have added 10 μl 2x buffer (my taq ready mix) 1μl...
05 May 2017 8,439 6 View
Hi everyone, I am trying to create a resistant cell line to a drug. Indeed, the first time i treated the cells with it , cells stopped growing then they grew again, when i passaged the cells to a...
04 April 2017 4,170 3 View
Dear all, I am giving this drug which is 16.5mM in DMSO and I want to make 0.1mM and 0.33mM of it to add it to the cancer cell line. I know DMSO might have an effect on the cells so I'm planning...
03 March 2017 5,566 4 View
Hi everyone I'm new to cell culture, so any help would be much appreciated. I need to replate 5X10^5 from 10cm dish into each well of 6 well plate. Any ideas of protocols to guid me? Thank you...
02 February 2017 4,162 3 View
what method/experiment can I use? Thank you
01 January 2017 9,205 3 View
Hi everyone, I am going to start my immunohistochemistry on monday and I was planning to make the PBS, 5% BSA in PBS, DAB, and ImmPRESS reagent on friday. Is it a good idea to make it before hand?...
01 January 2017 245 7 View
Hi everyone, I accidentally left the antibody on the bench for one hour. is that going to effect its binding ability? Thank you in advance for answering :)
01 January 2017 9,592 5 View
