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Questions related from Xianfeng Wang
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...
01 January 2017 4,596 9 View
How to prevent formamide being oxidized to formic acid? Thank you so much for any ideas.
11 November 2016 2,235 0 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...
03 March 2016 10,100 11 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...
03 March 2016 4,990 3 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...
10 October 2015 5,541 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...
05 May 2015 1,969 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...
01 January 2015 7,447 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...
09 September 2014 3,230 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...
05 May 2014 2,775 4 View
All freezers (-20C) we have are forest free. Is it safe to keep antibodies in those freezers? If safe, how many months?
03 March 2014 496 9 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...
03 March 2014 4,386 9 View
After the stimulation of macrophages with 2 drugs, I will collect cell culture medium and detect TNFa protein level with ELISA assay. What should be the best for ELISA value correction? Another...
10 October 2013 1,845 9 View
Usually there are several (even more than 10 or 20) antibodies for a protein in the market but how do you know which antibody is better or best for your experiment? Just try it one by one?...
07 July 2013 6,474 25 View
There are many kinds of BSA from Sigma. What BSA is good for blocking in phosph...
07 July 2013 4,236 5 View
I am preparing sodium pyrophosphate solution as a phosphatase inhibitor. I ordered both Na4P2O7 · 10H2O (Sigma S6422) and Na4P2O7 (Sigma P8010) from sigma and both are insoluble in water but it is...
06 June 2013 6,891 17 View
I am working with mouse Immunol cells. RAW264.7? Bone marrow derived macrophages? Peritoneal macrophages? or Spleen cells? or blood leukocytes?Thanks.
03 March 2013 3,155 0 View
