I need to measure RFP (red fluorescent protein) induction in FFPE sections. I know RFP is expressed by fluorescence when I dissect the brain, I just need to make some pretty-looking slides.
Cyclophosphamide is supposed to be activated by Cytochrome P450. I've treated Huh7 and PLC liver cancer cell lines with Cytochrome P450, but there is no sensitivity whatsoever, even at 500uM....
31 December 2016 1,025 6 View
I prepared a 50 mM phosphate buffer in H2O and pH'ed it to 7. I lyophilized it and dissolve it in D2O? What is the pD? And what would be the pT (I would like a tritium water solution, ultimately).
07 August 2016 2,244 2 View
I have an Emmons-Horner-like agent (triethylphosphonoacetate) that I would like to label with tritium. The alpha proton is readily exchangable in NaHCO3/D2O; I presume the same would work with...
07 August 2016 1,240 3 View
Many enzymes have preference for L versus D or R versus entiomers. Is there a precendent in the literature for changing, engineering by amino acid alteration, a D-selective into an L-selective enzyme?
02 March 2014 1,664 3 View
I have xenografted human tumor lysates that I am running western blots on. I need to determine the extent of mouse cell contamination in these lysates (I don't have slides for the tumors). I am...
05 June 2013 306 8 View
I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...
10 August 2024 7,480 3 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....
29 July 2024 5,007 0 View
I am doing immunohistochemistry of the c-Fos protein and I have a budget problem. For context, the experiment has been done over 3 years prior to my arrival and I have 40 brains done and 100 left....
29 July 2024 5,961 2 View
I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...
28 July 2024 366 3 View
I designed 2 dual hybridization probes for my single SNP. where I attach my fluorophore 5'-FAM-nnnn-BHQ1-3', and 5'-HEX-nnnnnn-BHQ2-3'. I need to run it on the MIC RT PCR machine by Biomolecular...
28 July 2024 5,658 0 View
I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...
28 July 2024 3,178 3 View
I'm having problems with ion exchange chromatography. After applying my sample (a fluorescent protein), a preferential path inevitably forms. The matrix I use is DEAE-Sephacel. Could someone...
27 July 2024 6,605 0 View
It has been long known that EDC carbodiimide and NHS easter's half life agains hydrolysis are highly related to pH. However, very surprisingly, not yet found a paper giving a table/chart of their...
25 July 2024 8,738 0 View