I would like to know what are CD markers for bovine DC.
I wish to make stable HEK293 cells with my gene of interest using pcDNA3.1 How can I proceed?
11 December 2014 8,919 17 View
i wish to express a protein eukaryotically in good quantity? which system i can go for? adeno viral vectors are good choice whether these vector have fusion tags to purify? (except yeast...
11 December 2014 9,711 2 View
I have cloned my protein into mammalian expression vector pVAC1 and subsequently transfected 6 ug of DNA into 293 cells using lipofectamine...protein will be secreted out of the cell, since vector...
09 October 2014 6,789 7 View
i have a protein that is not naturally secreted out of the cell..so i decided to add a secretory signal (eg. IL-2 signal sequence) at 5' end of the gene... kozak-secretory signal-my gene of...
09 October 2014 1,248 3 View
Can anyone suggest a protocol for extraction of nuclear and cytoplasmic proteins from cell line without kits and detergent solution (NP-40)... Can I use Nuclear Extract (NE) buffer (HEPES [20 mM]...
08 September 2014 5,971 7 View
I have precipitated cell culture supernatants with TCA (10%) followed by overnight incubation and acetone washing...then I have added Lammeli sample buffer, but I am unable dissolve the protein...
08 September 2014 2,061 17 View
I have cloned GFP at C-terminal of my protein to confirm its expression by GFP fusion After end of my protein 2 aminoacids are present before start of GFP I wanted to know whether this 2 a.a will...
07 August 2014 3,828 9 View
I am doing transfection of BHK 21 cells using Lipofectamine LTX plus. My vector is pVAX1, but I am not getting the expressed protein I included kozak sequence before my gene. The protocol which i...
02 March 2014 4,666 4 View
How to isolate monocytes from bone marrow? I need to further culture them for in vitro studies.
11 December 2013 3,458 5 View
I wish to express a recombinant protein that is not naturally secreted. How to signal a sequence so that it can be secreted out?
11 December 2013 4,933 1 View
Can anyone share any reference that gives mouse specific CD3 antibody sequence. I have been searching for this since few days and no luck. Thank you very much
08 August 2024 9,157 4 View
I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...
07 August 2024 7,535 4 View
I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...
07 August 2024 505 3 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
Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...
06 August 2024 641 2 View
i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...
05 August 2024 8,835 1 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
Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...
04 August 2024 8,894 1 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
I have used PE and PE-Dazzle 594 fluorochromes and have managed to get NK cells to properly show GranzymeB expression after 4 hr PMA/ionomycin stimulaton, but for some reason my CD8 cells in the...
01 August 2024 7,677 2 View
