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hey folks - I've recently picked up a new project, with very new techniques, and I've run into a minor stumbling block.
our collaborators have sent me some nanoparticle they have made. they have conjugated it to an antibody we use for targeting certain cell types. I need to assay for functionality to be certain our antibody targeting is not hindered by addition of the nanoparticle.
our typical plate-based cell binding assays involve adding our antibody to cells (human lymphoma and/or leukemia cells), going through a few wash steps, and assaying for remaining antibody (unbound Ab will be removed with supernatant when cells are pelleted).
well, the NP is very dense and can't be washed away. any spin that will pellet out cells will also pellet out Ab.
the NP is also magnetic, but I don't know a way to clearly pull the cells-Ab-NP complex out and quantify binding.
does anyone have any suggestions?
Topics I've Started
I've been off the radar for some time, but hope to be back a little more
27 February 2013 - 08:03 PM
WHEW, time flies!
in the past couple of years I haven't been on this forum much. my interests were wandering in other directions for awhile, but I'd like to spend time hear again, and at least get a feel for all the changes. for now, I'd like to start with a (re-)introduction?
when I participated heavily in this forum several years ago, I was primarily a molecular biologist. I did fluorescent staining of cells, cloned and expressed proteins, working primarily with small peptides and DNA. I ran a ton of qRT-PCR. I worked a little with primary cells.
now, I do leukemia research, primarily on mice, as part of a very large group. I work a lot with radiolabeled antibodies and am getting into nanoparticles, and I do a lot of flow cytometry. I work with a ton of heme cancer cell lines and have a large number of disease models for leukemia, lymphoma, and other blood cancers.
as far as background, I have 2 BS (one in Microbiology; one in Molecular Biology and Biochemistry). I have also been a Public Health Microbiologist, and did a load of cloning (and everything else DNA) back in the late 90's. so, I'm rusty with those techniques, but do have a solid foundation
back in the day I was a Mod, and while I'm still on the list I've PM'd our gracious, intrepid SuperAdmin to see if I'm still officially on the team after my absence, lol.
for those of you who used to know me...Hi! for those of you who have no clue why I'm rambling on...Hi!
A
in the past couple of years I haven't been on this forum much. my interests were wandering in other directions for awhile, but I'd like to spend time hear again, and at least get a feel for all the changes. for now, I'd like to start with a (re-)introduction?
when I participated heavily in this forum several years ago, I was primarily a molecular biologist. I did fluorescent staining of cells, cloned and expressed proteins, working primarily with small peptides and DNA. I ran a ton of qRT-PCR. I worked a little with primary cells.
now, I do leukemia research, primarily on mice, as part of a very large group. I work a lot with radiolabeled antibodies and am getting into nanoparticles, and I do a lot of flow cytometry. I work with a ton of heme cancer cell lines and have a large number of disease models for leukemia, lymphoma, and other blood cancers.
as far as background, I have 2 BS (one in Microbiology; one in Molecular Biology and Biochemistry). I have also been a Public Health Microbiologist, and did a load of cloning (and everything else DNA) back in the late 90's. so, I'm rusty with those techniques, but do have a solid foundation
back in the day I was a Mod, and while I'm still on the list I've PM'd our gracious, intrepid SuperAdmin to see if I'm still officially on the team after my absence, lol.
for those of you who used to know me...Hi! for those of you who have no clue why I'm rambling on...Hi!
A
