18 Questions 28 Answers 0 Followers
Questions related from Xingchen Dong
Dear all, I treated wild-type cells and knockout cells with two drugs X and Y for 24 hours. Then I measured the Gene A expression. Drug Y significantly increased Gene A expression around 500-fold...
14 March 2024 1,262 0 View
Hi, I am wondering if anyone could share some thoughts on my qRT-PCR results. I sonicated the mouse intestine (homogenizer would be better?) and extracted the total RNA with Trizol. Then I...
01 November 2022 5,486 6 View
Hi, I am wondering if anyone is familiar with Biorad Trans-blot turbo transfer system. When we purchased this machine, they gave us a 5x buffer, paper stack, and PVDF membrane kit. Indeed, it...
15 April 2022 5,510 3 View
Dear all, I am curious that if anyone knows whether surface markers such as F4/80, or CD206 are stable in paraffin sections? The small intestine paraffin sections are about 3 years old, and I...
14 August 2021 8,009 3 View
Hi all, I am curious that why in some methods, people add IL1b and IL23 on top of the TGFB and IL6 for Th17 cells differentiation in vitro? Also, the percentage of Th17 after in vitro...
19 May 2021 821 4 View
Dear all, I used to use L929 conditional medium for BMDM isolation, and I generally get around 100 million BMDMs per mouse (8-12 weeks) after 7 days of differentiation (10 Petri dishes with ~10...
26 March 2021 3,422 8 View
Dear all, I've been working with the bone marrow-derived macrophages for a long time and now I am trying to culture the mouse macrophage cell line, the Raw264.7 cell line. I used the Gibco DMEM,...
05 March 2021 7,494 2 View
Hi all, Our lab is trying to activate pyrin inflammasome in bone marrow-derived macrophages with TcdB toxin from Clostridium difficile. We are not a microbiology lab and we have to purchase the...
28 July 2020 5,374 1 View
Hi guys, I am a little bit confused by the concept that priming BMDMs with LPS for Nlrc4 inflammasome studies. Originally I thought this LPS priming is essential for Nlrp3 due to it activates...
03 November 2019 403 4 View
Hey guys, I am trying to PCR out 0 to -1k region of three genes. I used pfu-ultra and followed strictly the protocol. I adjusted everything to their proposed working concentration. On top of...
14 May 2019 2,957 0 View
Hi Guys, I new to immunology and I have a simple question about LPS stimulation on BMDMs, also known as 'LPS priming'. I read something that says LPS binding to TLR4 needs LPS binding...
01 May 2019 9,663 9 View
Hi guys, I tried to transfect poly dA:dT into macrophages to activate Aim2 inflammasome. I setup 4 wells, the first two of which have lps and transfection reagent as negative control. The other 2...
20 September 2018 5,994 5 View
Hi guys, I am trying to see inflammasome activation after delivering flagellin from salmonella into macrophages. I use DOTAP transfection reagent from Roche which is widely used by other...
15 August 2018 4,593 4 View
Hi guys, Using the PDB NGL viewer, I can see there is a ligand (such as a drug inhibitor) binding to a pocket of my interest protein. But when i download this PDB file and visualize it with VMD,...
15 December 2017 2,031 3 View
Hi guys, Where can I find the gene expression level in multiple cell lines, such as I want to see the level of gene A in multiple gastric cancer cell lines? And this level could be either mRNA or...
16 August 2017 4,151 9 View
Hi guys, Is the MPO activity equivalent to the neutrophil number? I mean if I observed an increase in MPO activity, can I assume that there's an increase in neutrophil number?
07 June 2017 7,663 4 View
Hi guys, I am trying to quantify the miRNA by using real-time PCR. After the RNA extraction, I need to add the poly A tails to mature miRNAs and I would assume that a poly A tailing kit should be...
01 July 2016 1,969 3 View
Hi guys, I am trying to insert a 20bp fragment into a vector regardless of the directionality of the fragment. The enzymatic sites I picked are XhoI and MluI. Therefore I added these two sites...
01 January 1970 6,731 2 View
