I am working with mouse Immunol cells. RAW264.7? Bone marrow derived macrophages? Peritoneal macrophages? or Spleen cells? or blood leukocytes?Thanks.
I am doing a time course experiment. Treat cells with a drug for different times (0h, 2h, 8h, 18h, and 24h, respectively), and then harvest cells for RNA to check cell responses. So here we have 2...
31 December 2016 4,650 9 View
How to prevent formamide being oxidized to formic acid? Thank you so much for any ideas.
10 November 2016 2,286 0 View
I treated cells with stearic acid for different times. Collect total protein lysate after 3-time wash with PBS. Test protein concentration with Bio-Rad DC Protein Assay Reagents Package...
02 March 2016 5,041 3 View
Usually people add M-CSF or L929 conditioned medium to regular medium (e.g. DMEM + 10% FBS) to differentiate bone marrow into BMDM (bone marrow derived macrophages) for 7 days. I have an argument...
02 March 2016 10,163 11 View
I am trying to co-transfect 3 plasmids into macrophage cell line (RAW264.7). I can transfect single plasmid successfully and can detect with immunoblotting. But it is very hard for me to do...
09 October 2015 5,579 8 View
Up till now, there are two kinds of 2x Laemmli sample buffers: Buffer 1) 65.8 mM Tris-HCl, pH 6.8, 2.1% SDS, 26.3% (w/v) glycerol, 0.01% bromophenol blue Please refer to the original...
04 May 2015 2,015 8 View
I treated macrophage cells with LPS (1h to 24h time course) and tried to detect p65 acetylation. Method 1) I collected total cell lysate with RIPA buffer containing 10mM NAM and 10uM TSA. And then...
31 December 2014 7,494 8 View
We have a mouse hybridoma cell line which can secrete a mouse IgG2a monoclonal antibody. The culture medium is RPMI 1640 medium + 10% horse serum + NEAA +1 mM sodium pyruvate + penicillin (100...
08 September 2014 3,274 4 View
I am working with HUVEC cells and hope to detect E-Selectin, VCAM-1 and ICAM-1 protein expression with western blot and cell ELISA (for surface expression). Does anybody have experiences with...
04 May 2014 2,832 4 View
Here is my protocol: 1. Collect small intestine from mice and push out the stool. 2. Flash freeze in liquid nitrogen and then store the tissue at -80C for later use. 3. Homogenize tissues in RIPA...
02 March 2014 4,438 9 View
I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...
07 August 2024 6,034 3 View
After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...
05 August 2024 9,637 2 View
Brain and body mass together are positively correlated with lifespan (Hofman 1993). The duration of neural development is one of the best predictors of brain size, and conception is the best...
05 August 2024 6,247 3 View
I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...
04 August 2024 9,913 7 View
Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...
04 August 2024 7,282 2 View
In my experiment, I treated RAW macrophages with 100 ng/mL of LPS. After 24 hours, I collected the supernatant and replaced it with fresh media. I then quantified the cytokine levels of TNF and...
01 August 2024 476 2 View
Some Staphylococcus aureus strains Inhibit the growth of Mycobacteria in Mueller Hinton Agar medium containing 10% OADC. Do some Staphylococcus aureus strains have in vitro antimycobacterial activity?
29 July 2024 10,023 2 View
I have a question about human macrophages cell surface markers.I want to stain the unpolarized human macrophages for flow cytometric sorting without permeabilizing them. It is because I need to...
29 July 2024 236 2 View
I created a file with my outgroup and ingroup species using Beauti, ran it in BEAST, viewed it in Tracer, and then used TreeAnnotator to create a file that I imported into RASP. Could someone...
28 July 2024 2,979 1 View
PeptiCloud (www.pepticloud.com) is a bioinformatic platform that allows researchers to organize and share their data for their projects as well as collaborate with others in one place. Through...
28 July 2024 4,762 2 View