meta user
network profile
| bio | website | pigimal.com |
|---|---|---|
| location | Massachusetts | |
| age | 34 | |
| visits | member for | 11 months |
| seen | 20 secs ago | |
| stats | profile views | 9 |
Long-time OSS user/admin, beginning serious coder - I heart Python. Master of Science in Molecular Medicine, many years' experience in biotech working with antibodies and related assays. Science and computer geek. Dad.
|
54m |
comment |
How difficult is to renature a protein? @shigeta - I was specifically thinking of antibodies (150 kDa, 4 subunits disulfide-bonded to each other). At my previous job we experimented with purifying antibodies by eluting them from an antigen column with varying concentrations of guanidine, then dialyzing directly into PBS. We didn't do any biophysical characterization to look at the extent of denaturation, but in our functional assays they performed similarly to those eluted by the standard (pH-dependent) method. |
|
2d |
comment |
Writing a paper Tag should probably be Taq... |
|
2d |
comment |
Writing a paper You should change your "Amplification..." sentence to remove the word "condition" and change 25 ml to 25 µL, but other than that it looks okay. Also, maybe spell out "Forward" and "Reverse" when listing your primers. |
|
May 16 |
comment |
What material is recommended for autoradiography cassettes when working with 32P? If you're worried, contact your local radiation safety committee - they should have a wide-spectrum detector around that can detect Bremsstrahlung. I've only ever used aluminum/plastic cassettes myself for autorads. The phosphor screen should block a good deal, but you do not want to take any chances where radiation is concerned... |
|
May 13 |
comment |
SNPs mapping into protein thank you for using one of my favorite gene names :) |
|
May 13 |
comment |
SNPs mapping into protein I assume SNP means Single-Nucleotide Polymorphism? I'm confused exactly what your question is - how does one take a SNP and see what effect it has on the protein sequence? Is that it? |
|
May 11 |
comment |
5-HT2A Receptor I'll answer more thoroughly if you edit and reopen the question, but in the meantime I'd suggest checking out PyMol, a great protein (and other structure) visualizing tool. |
|
May 7 |
comment |
Are there more descriptive ways of naming genes and gene interactions? there are actually MAP4Ks as well - Mitogen Activated Protein Kinase Kinase Kinase Kinases :) |
|
Apr 25 |
comment |
Up to date B cell review Do you have any references for the BCR cross linking model dispute? I'm not working directly in the field now, so I'm not as up to date as perhaps I should be... |
|
Apr 25 |
comment |
Up to date B cell review @AndroidPenguin If you're looking for reviews on specific subjects in journals, some of the journals I turn to include Annual Reviews in Immunology, Trends in Biochemical Sciences, Cell, Nature Reviews, Nature Reviews Immunology, and Trends in Immunology, among others. It's quite possible to find great reviews in lower-impact journals like the Journal of Immunology, and you'll often find more esoteric reviews there as well, but for now I'd stick with the real biggies like the ones I mentioned. |
|
Apr 24 |
comment |
Up to date B cell review what specifically about B cells are you interested in? The possible subject matter is so broad that I doubt you'll find reviews covering all of B cell biology and function. A recent immunology text might be better. |
|
Apr 24 |
comment |
Role of Fbx15 in ES cells and its use in assaying for iPS cells (Yamanaka paper and others)? @dd3 I don't know if Fbx15 is so much a marker of iPS cells as it is just a target of the Oct4 transcription factor, meaning its expression will be active in Oct4+ cells, providing a convenient way of driving exogenous (and then endogenous) expression of the Yamanaka factors long-term. |
|
Apr 15 |
comment |
What is the difference between HPLC and FPLC and why is FPLC preferable for protein purification? Unfortunately, several of your points are just wrong. Anything an FPLC can do, an HPLC can do, but an HPLC can also do more. An FPLC is essentially a specialized HPLC for proteins. High pressure is not required in HPLC, but it is an option, depending on the type of column. I assume by "solvent" you mean organic solvents, but that is incorrect - an HPLC can use a salt/buffer mobile phase just as easily as an FPLC. I use vertical columns in an HPLC all the time. Temperature doesn't have to matter in an HPLC, it's all in the parameters you set for a particular experiment. |
|
Apr 13 |
comment |
How does Tianeptine work to elicit its therapeutic effects? so what's wrong with these theories? Wikipedia has some links to others if you're interested... |
|
Apr 12 |
comment |
What would be the best design for spike-and-recovery and linearity-of-dilution validation experiments in one 96-well ELISA plate? I think that looks like a good start. You may need to adjust your serial dilution factor so that you are completely within the linear range of your standards. I would suggest that you think very hard about assay vs. sample buffer, perhaps doing a titration experiment to compare their performance. Any way you look at it, it will be impossible to do a complete validation with one plate, since there are too many variables to test. As long as you have your standard curve, blanks, and at least one common sample on each plate, you'll be able to compare your data across plates/runs. |
|
Apr 12 |
comment |
What would be the best design for spike-and-recovery and linearity-of-dilution validation experiments in one 96-well ELISA plate? you now have 11 points, please post any relevant images |
|
Apr 10 |
comment |
Where to put the gene after eukaryotic promoter for best expression levels? Take a look at the sequences for some common/popular vectors for your target organism. As WYSIWYG pointed out, it doesn't need to be very big, and it can vary somewhat depending on where in the multiple cloning site you insert your gene of interest. |
|
Apr 9 |
comment |
Iron deficiency anemia symptoms explanation @WYSIWYG I'm not sure what collagen has to do with it - fingernails and hair are make up primarily of keratins... |
|
Apr 9 |
comment |
Iron deficiency anemia symptoms explanation sorry, I could have worded it better, I was in a hurry :) Yes, anemia (in general) is defined as a deficiency in something, and in common usage the word anemia alone is typically taken to mean iron anemia or iron deficiency. What I meant is that copper levels can also be affected by iron anemia. |
|
Apr 8 |
comment |
Can I heat Trizol? As far as I know, Trizol disrupts protein-RNA complexes on its own - check the protocol. Also, phenol, a major component, is flammable. I don't know what the flash point of Trizol is, but I would be extremely careful. This protocol claims you can heat it. Does your experiment not work at RT? |
