17 Questions 45 Answers 0 Followers
Questions related from Doaa Glal
I have KO cancer cell line, and I am applying shRNA to target some genes. My control is RFP-shRNA. I ran western blot to check my protein of interest, and i have found that KO cell line with...
09 September 2018 5,238 6 View
Hello everyone, I have unexplained phenomenon in my data regarding one cytokine. The level of this cytokine by Qpcr is very high in knockout more than WT mice in naive condition and after...
09 September 2018 4,531 3 View
Dear fellow, I am planning to send samples of cancer cell lines to NGS. I have one set unstimulated, and one set with cytokine stimulated. However, I am not sure if it is better to stimulate...
06 June 2018 10,123 2 View
Hello fellows, I am doing EMSA. I wanted to check for the binding and efficacy of DNA and nuclear extract proteins. So, I run Acrylamide gel (native, 5% , 0.5xTBE buffer, with DNA view dye )....
04 April 2018 8,081 23 View
Hello, I'm doing EMSA using adherent cell line . I extracted the nucleus part but it was very viscous. Why the nuclear extract was very viscous ? And can I apply sonication ? Thank you.
04 April 2018 8,967 4 View
Dear bacteria and cell culture specialists, Recently we encountered this bacteria during cell culture, please see the attached images. These bacteria doesn't change the PH of the culture media...
04 April 2018 2,625 4 View
Hello fellows, How can I find a promoter sequence for a certain gene ? ( simple way to explain , i don't have much bioinformatic background) Thanks in advance
03 March 2018 9,811 11 View
Hello guys, I am running western for the 3rd time for the same protein. First two times I can see clear one band. It is inducible protein. However, on the 3rd time, I have two bands , instead...
02 February 2018 2,729 2 View
Hello Fellows, I want to clone a protein with a Flag but I can't decide where should I place the Tag on C-terminal or N-terminal since some papers put it on N-terminal but others put on...
11 November 2017 1,519 4 View
Dear fellows, I am facing a problem using Isotype control, for example, when I run flow , the signal in un-stimulated samples is 10%, and in stimulated samples is 12%. However, the isotype...
11 November 2017 8,528 2 View
Hello Guys, Can someone provide me with a simple protocol to measure intracellular calcium ? I want to isolate intestinal epithelial cells from mice and measure Calcium level ? Can we detect...
08 August 2017 1,063 6 View
Hello, I want to use Tamoxifen to induce cre-knockout mice in Jacson lab, they mentioned to dissolve tamoxifen in corn oil at a concentration of 20 mg/ml by shaking overnight at...
07 July 2017 7,730 2 View
Hello Fellows, My questions is regarding Qpcr. When I run Qpcr , I do replicates from the same sample to make sure data is consistent. House Keeping gene (L32) is always very consistent with me in...
05 May 2017 6,188 4 View
Hi fellows , I want to know How long can I store the cell line in -80C before liquid nitrogen? Thanks.
01 January 2017 7,089 7 View
Hello, I'm new for flow cytometry. I have purchased Rabbit monoclonal AB which I usually use for immunofluorescence staining. Now, I want to use it for intracellular flow cytometry staining. How...
11 November 2016 5,247 7 View
Hello, I am infecting mice orally with salmonella 1344. I used two doses 10e4 and 10e6. I did fecal CFU on day 3 and day 5 post infection to detect salmonella titre in feces. However, no colonies...
03 March 2016 1,230 3 View
I infected different mice with bacteria with the following dose titer 10e9. 10e8, and 10e7. On day 7, I took the feces and did serial dilutions from 10e1-10e8 dilution. However, the colonies...
11 November 2015 4,589 3 View
