Re: [ccp4bb] anti-His6 Ab
http://www.mail-archive.com/ccp4bb@jiscmail.ac.uk/msg27397.html Since the discussion, we tried qiagen ab, and I was quite happy with it. I did not do extensive performance tests, but it was better than the previous ab. Regards, Juha On Friday, 15 March 2013, Elias Fernandez wrote: Dear All, would you be able to recommend a primary anti-His6 Ab for western blotting? Thus far, the ones we’ve used are not very specific to the N-terminal hexahistidines and we pick up lots of background. Also, there’s one (others?) from Sigma that is conjugated to HRP, but costs ~$650. Thanks! Elias -- Genius may have its limitations, but stupidity is not thus handicapped. -Elbert Hubbard Juha Vahokoski Kalliotie 16 as. 10 90500 Oulu Finland mobile:+358 40 5286 778
Re: [ccp4bb] Off-topic His-Antibody
I have been using an HRP conjugated antibody from Thermo that can detect maybe 3-8ng of HisTagged protein from insect cell lysates. Performance is ok, but with the sensitivity I am not so happy with. How is the sensitivity with the Qiagen antibody? How many picograms you need for a clear band? Does anyone know a good HRP conjugated antibody? Regards, Juha On 26 June 2012 06:59, Luca Jovine luca.jov...@ki.se wrote: We routinely use BSA-free anti-5His mAb from QIAGEN, and it works just fine (at least using media and lysates of both insect and mammalian cells): http://www.qiagen.com/products/protein/detection/qiaexpressdetectionsystem/penta-hisantibody.aspx Good luck, Luca Luca Jovine, Ph.D. Assistant Professor EMBO Young Investigator Karolinska Institutet Department of Biosciences and Nutrition Center for Biosciences Hälsovägen 7, SE-141 83 Huddinge, Sweden Voice: +46.(0)8.524-81136 FAX: +46.(0)8.6081-501 E-mail: luca.jov...@ki.se W3: http://jovinelab.org From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of D Bonsor [ dbon...@ihv.umaryland.edu] Sent: Monday, June 25, 2012 23:33 To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Off-topic His-Antibody Are there any good antibodies for His-tags on the market. I have never used one and I heard several stories that they were poor and not worth the money, over the past few years. The only post I could find on the BB was from 2008. Have His-Antibodies improved or are they still rubbish. -- Genius may have its limitations, but stupidity is not thus handicapped. -Elbert Hubbard *
Re: [ccp4bb] detect dsDNA
Also this method might be useful if crystals tolerate the treatment. Regards, Juha Fluorescence detection of nucleic acids and proteins in multi-component crystalshttp://onlinelibrary.wiley.com/doi/10.1107/S0907444905035365/abstractActa Crystallographica Section D Volume 62, Issue 2, February 2006, Pages: 146–150, Hubert Kettenberger and Patrick Cramer DOI: 10.1107/S0907444905035365 doi:%C2%A010.1107/S0907444905035365 On 1 October 2011 06:36, zq deng dengzq1...@gmail.com wrote: Hi all, . recently,I got a crystal of protein-DNA crystal.i used silver stainto prove that it is a protein crystal.Does anyone have method to detect if there is DNA in the crystal. any suggestion will be appreciated. Regards, deng
Re: [ccp4bb] Off topic: vector map editing and DNA sequence alignment software
Dear CCP4Pers, also j5 might be a good choice. I am not personally familiar with the software, so I would happy to hear any comments. It also seems to cope with modern cloning techniques like SLIC. clicking away at http://j5.jbei.org/index.php/Main_Page Regards, Juha On 27 September 2011 18:42, Florian Schmitzberger schmitzber...@crystal.harvard.edu wrote: Dear All, What type of software are people commonly using these days for vector/plasmid map editing, making/visualizing vector maps, and aligning (small to medium size) DNA sequencing data? Preferably, it should not be too expensive and be able to write text files, readable by other programs. I am familiar with VectorNTI, which is great for vector visualization and editing; but I find it somewhat expensive. Sequencher seems good to quickly align DNA sequences (such as from DNA sequencing) with templates, but is not free. I have been using ApE for while for alignments, but aligning many sequences is more cumbersome than in Sequencher; I have not tested if Sequencher is good at visualizing and editing plasmid maps. Ideally, I would like to have a single program for both purposes (vector editing and DNA sequence comparison). Does something like that exist? What are the alternatives to above programs? Thank you in advance. Florian --- Florian Schmitzberger, PhD Biological Chemistry and Molecular Pharmacology Harvard Medical School 250 Longwood Avenue, Seeley G. Mudd 123 Boston, MA 02115, US Tel: 001 617 432 5603 -- Genius may have its limitations, but stupidity is not thus handicapped. -Elbert Hubbard Juha Vahokoski Kalliotie 16 as. 10 90500 Oulu Finland mobile:+358 40 5286 778
Re: [ccp4bb] Nucplot -where to download
Dear Joyce, PDBsum/EBI also produces Nucplot output. You can also upload you own file for analysis. also this might help you http://monster.northwestern.edu/ If you find any other alternatives, I am happy to hear them. HTH, Juha On 5 November 2010 21:06, Joyce wrou...@yahoo.com wrote: Hi, I have been trying to download the program nucplot from http://www.biochem.ucl.ac.uk/bsm/nucplot/avail.html but the files are no longer available there. Could somebody tell me where I can download the program, or if there is an equivalent program I could use to make a schematic of protein-DNA interactions? Thank you, Joyce
