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[ccp4bb] Defining consensus pattern for searching a motif in protein database(s)
Dear All I want to search SUMO interacting motif (SIM) in different protein data bases and also in the given protein sequences. Does anyone can provide me the PROSITE AC and/or ID for the SIM motif? I will be grateful if you provide me MATRIX for the SIM motif profile. I do not know much about how to define the amino acid sequence ‘pattern’ for searching the motif in the ‘Prosite’ tool. For an example AP endonuclease contains following consensus pattern D-[ST]-[FY]-[RP]-[KHQ]-x(7,8)-[FYWD]-[ST]-[FYW] (2) What is the meaning of x(7,8) and (2) in the above example. Any literature or article regarding this topic is most welcome. Thank you in advance Raj E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile)
Re: [ccp4bb] how to quantitate protein which dont have ne aromatic residue
Hi Arpita You can try QUANTI-iT Protein assay kit from Invitrogen. But still there is nearly 20-50% discrepancy between this method and a Abosorbance at 280. I also faced same problem with a protein, then re-cloned by adding a Trp at the C-terminus. Raj E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) --- On Sat, 9/4/11, Arpit Mishra ar...@igib.in wrote: From: Arpit Mishra ar...@igib.in Subject: [ccp4bb] how to quantitate protein which dont have ne aromatic residue To: CCP4BB@JISCMAIL.AC.UK Date: Saturday, 9 April, 2011, 3:22 PM hello everybody i am working on the protien which dont have any aromatic residue i do fplc other purification using 220 absorption, but i want to quantitate protein precisely i have tried using BCA nd bradford but both methods quantification is not matching,,so any one is having sum idea how to quantitate it precisely thanks in advance for your valuable suggestion.. Arpit Mishra
[ccp4bb] Question on calculation of RMSD
Dear All I have two structures of homo-dimeric protein complex with different DNA. I want to calculate RMS deviation between second monomer from these two complexes by fixing superposed first monomer. This I require to know what is the effect of DNA on relative orientation of two monomers in the dimer. Previously I was using MOLEMAN2 to do this calculation. Please can you suggest me any other program to do this calculation. Thanking you Raj E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile)
[ccp4bb] Protein crystallizes while concentration
Dear All I am Rajkumar, working on the protein which has unusual behavior while concentration. When I kept protein in 15 mM Tris 7.5, 150 mM NaCl and 5 mM DTT, the solubility of the protein is decreases drastically and tend to crystallize while concentration. Protein cannot be concentrated more than 3 mg/mL, however I noticed white turbid protein if I force to concentrate 3mg/mL. When I observed this white turbid solution under the microscope, I noticed shower of tiny protein crystals which are needle in shape. I screened freshly purified protein (2.5 mg/mL) in different Hampton and Qiagen screens, strangely none of the conditions gave the crystals. I concentrated left over protein at 15oC at 3 mg/mL and kept in the 4oC for 4 days again I noticed shower of crystals. This protein solubility is increased to ~20mg/mL when I kept in 15 Tris 7.5, 400 mM NaCl, 5% Glycerol and 5 mM DTT, but did not crystallize while concentration and also after screening with Hampton and Qiagen screens. My queries are 1. How do I get the crystals in the crystallization set up rather than while concentration, so that I can control the diffusion and finally nucleation? 2. Could anybody give me suggestions on seeding in this type of situation? 3. Any comments on reverse vapor diffusion for this type of protein are most welcome. So I can keep protein in high ionic strength (~400 mM NaCl)and diffuse against low Ionic strength or deionized water? Or any other protocol? Any suggestions are well appreciated. Thanking you in advance Raj E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your new Email address! Grab the Email name you#39;ve always wanted before someone else does! http://mail.promotions.yahoo.com/newdomains/aa/
[ccp4bb] Error message while refining protein-DNA complex structure in Refmac5
Dear Garib Thank you. After removing SCALE card in the pdb file, refinement is completed somthly. Rajakumara --- Garib Murshudov ga...@ysbl.york.ac.uk wrote: It seems that something may be wrong with your input file. Specifically with the SCALE card in the pdb file. Could you please remove SCALE lines from the pdb and try again. If it does not help then could you please send me your pdb file and I will try to sort out. Garib On 16 Mar 2009, at 22:43, E rajakumar wrote: Dear All I am refining proitein-DNA complex structure in Refamac5. When I used coordinate file containing 2 bases less, then the refinement is running smoth and perfect. But when I built 2 exta bases to the existing DNA in the coot then refinement is failed with the following error message. /usr/local/ccp4-6.1.0/bin/refmac5 XYZIN/usr6/rajkumar/APS/hem/mar9/H2/molrep/BCDNA-built2-NCS- refm2.pdb XYZOUT /tmp/rajkumar/hemCG_19_2_pdb_1.tmp HKLIN /usr6/rajkumar/APS/hem/mar9/H2/molrep/P6122.mtz HKLOUT /tmp/rajkumar/hemCG_19_3_mtz_1.tmp LIBOUT /usr6/rajkumar/APS/hem/mar9/H2/molrep/hemCG_19_lib.cif has failed with error message At line 2486 of file /usr/local/xtal/ccp4-6.1.0/src/refmac5_/make_PDB.f Fortran runtime error: Bad value during floating point read It seems there is error in LIB file generation. Coordinate format and atom labelling is accoring to refmac convention. Please can anybody suggest me how do I trooubleshoot. Thanking you Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your preferred Email name! Now you can @ymail.com and @rocketmail.com. http://mail.promotions.yahoo.com/newdomains/aa/ E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your preferred Email name! Now you can @ymail.com and @rocketmail.com. http://mail.promotions.yahoo.com/newdomains/aa/
Re: [ccp4bb] Error message while refining protein-DNA complex structure in Refmac5
Dear Garib Thank you very much as you suggested, I removed SCALE card in the pdb file, then it is running smothly. Rajakumara --- Garib Murshudov ga...@ysbl.york.ac.uk wrote: It seems that something may be wrong with your input file. Specifically with the SCALE card in the pdb file. Could you please remove SCALE lines from the pdb and try again. If it does not help then could you please send me your pdb file and I will try to sort out. Garib On 16 Mar 2009, at 22:43, E rajakumar wrote: Dear All I am refining proitein-DNA complex structure in Refamac5. When I used coordinate file containing 2 bases less, then the refinement is running smoth and perfect. But when I built 2 exta bases to the existing DNA in the coot then refinement is failed with the following error message. /usr/local/ccp4-6.1.0/bin/refmac5 XYZIN/usr6/rajkumar/APS/hem/mar9/H2/molrep/BCDNA-built2-NCS- refm2.pdb XYZOUT /tmp/rajkumar/hemCG_19_2_pdb_1.tmp HKLIN /usr6/rajkumar/APS/hem/mar9/H2/molrep/P6122.mtz HKLOUT /tmp/rajkumar/hemCG_19_3_mtz_1.tmp LIBOUT /usr6/rajkumar/APS/hem/mar9/H2/molrep/hemCG_19_lib.cif has failed with error message At line 2486 of file /usr/local/xtal/ccp4-6.1.0/src/refmac5_/make_PDB.f Fortran runtime error: Bad value during floating point read It seems there is error in LIB file generation. Coordinate format and atom labelling is accoring to refmac convention. Please can anybody suggest me how do I trooubleshoot. Thanking you Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your preferred Email name! Now you can @ymail.com and @rocketmail.com. http://mail.promotions.yahoo.com/newdomains/aa/ E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your new Email address! Grab the Email name you#39;ve always wanted before someone else does! http://mail.promotions.yahoo.com/newdomains/aa/
[ccp4bb] Error message while refining protein-DNA complex structure in Refmac5
Dear All I am refining proitein-DNA complex structure in Refamac5. When I used coordinate file containing 2 bases less, then the refinement is running smoth and perfect. But when I built 2 exta bases to the existing DNA in the coot then refinement is failed with the following error message. /usr/local/ccp4-6.1.0/bin/refmac5 XYZIN/usr6/rajkumar/APS/hem/mar9/H2/molrep/BCDNA-built2-NCS-refm2.pdb XYZOUT /tmp/rajkumar/hemCG_19_2_pdb_1.tmp HKLIN /usr6/rajkumar/APS/hem/mar9/H2/molrep/P6122.mtz HKLOUT /tmp/rajkumar/hemCG_19_3_mtz_1.tmp LIBOUT /usr6/rajkumar/APS/hem/mar9/H2/molrep/hemCG_19_lib.cif has failed with error message At line 2486 of file /usr/local/xtal/ccp4-6.1.0/src/refmac5_/make_PDB.f Fortran runtime error: Bad value during floating point read It seems there is error in LIB file generation. Coordinate format and atom labelling is accoring to refmac convention. Please can anybody suggest me how do I trooubleshoot. Thanking you Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your preferred Email name! Now you can @ymail.com and @rocketmail.com. http://mail.promotions.yahoo.com/newdomains/aa/
[ccp4bb] Cryoprotectant for protein-DNA complex crystal
Dear All I am working on protein-DNA complex crystals for data collection. These crystals are grown in 15-20 % of PEG3350 or PEG4000 with pH of 6 to 7. When I soak the crystals more than a minute in the cryo solution (15-20% of Glycerol or ethylenglycol + reservoir) the resolution of diffraction is becoming weak (reducing to 6.0 A from 4.5 A) and also the spots are getting spread (increase in mosaicity). Appears that Glycerol or Ethylene glycol not good cryoprotectants in this case. Is there any study on effect of cryoprotectant on protein-DNA complex crystal and protein-DNA complex dissociation? I also want to know which type (organics, oils, polyols, sugars, polymers ) of cryoprotectant is most preferred in protein-DNA complex crystal. Thanking you in advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your new Email address! Grab the Email name you#39;ve always wanted before someone else does! http://mail.promotions.yahoo.com/newdomains/aa/
[ccp4bb] Protein-DNA complex prepartion for crystallization
Dear All Sorry for non-crystallography query. I am working on DNA binding protein, while mixing DNA with protein for preparing Protein-DNA complex for crystallization, protein is precipitating. pI of the protein is 9.3 and in 15 mM HEPES 7.0, 150mM NaCl and 5% glycerol. Concentration of the protein used for mixing with DNA is 8 mg/mL. DNA to protein molar ratio is 1.2. Please advise me how to prevent precipitation. Is changing of buffer pH and adding divalent cation like MgCl2 can help in preventing precipitation? Thanking You Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your preferred Email name! Now you can @ymail.com and @rocketmail.com. http://mail.promotions.yahoo.com/newdomains/aa/
[ccp4bb] Query on program for creating random crystallization screen
Dear All I want to create a random crystallization screen using given set of crystallization parameters (pH, Precipitant, salt, additive.) for protein-DNA complex crystallization. I was using Brent Segelke's CRYSTOOL program, not accessible online now. Please can you suggest any other such programs and/ how do I can access the CRYSTOOL (sort of registration or purchase?) Thanking you in advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your preferred Email name! Now you can @ymail.com and @rocketmail.com. http://mail.promotions.yahoo.com/newdomains/aa/
[ccp4bb] Ammonium citrate tribasic buffer
Dear All Sorry for non crystallographic query. Can any body mail me how to prepare Ammonium citrate tribasic (citric acid triammonium salt) buffer pH 6.7 to 7.25 and also what is the pKa value. Thanking you in advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Get your new Email address! Grab the Email name you#39;ve always wanted before someone else does! http://mail.promotions.yahoo.com/newdomains/aa/
Re: [ccp4bb] query on DNA-protein complex preparation for crystallization
Hi Artem Evdokimov Thank you for the mail. I have synthesized DMT-on oligos in our laboratory. Deprotection was performed treating with ammonium hydroxide for 15 hours at 55C. Then, DMT-on oligo was separated from off using RPHPLC. DMT was cleaved by treating with 20% glacial acetic acid for one hour. Then, DMT-off DNA was separated from DMT, again using RPHPLC. Lyophilized DMT-off oligos were dissolved in 3 mL of Milli Q water and dialysed against 2 L of milli Q water for 4hrs by changing water 2 times. Then complemntary oligos are concentrated around 1.0 mM and mixed them and concentrated further to 1.5 mM (duplex). 2 mM (final concentration) of Magensium chloride was added to oligos and concentrated to half of the volume. While concentrating oligos become viscos and white precipitate. however, annealing did not help to dissolve the white precipitate. I kept oligos in distilled water, without adusting pH. Please can you mail if I iginite DNA on metal spatual, eiether burns or not, what it indicates? Thanking you Rajakumara [EMAIL PROTECTED] wrote: Hi, How did you synthesize the DNA? I assume external vendor (so few people make their own these days)? How was the DNA purified? Sometimes if only a 'desalting' step is used there may be 'other chemicals' in the mix. Also, what pH was your DNA at, and in what buffer (if any)? If your DNA degraded you may have Pi in solution, which forms insoluble precipitates with many counterions. So, first of all I would check your white precipitate - does it dissolve in anything at all? If it does dissolve, what pH does it have? Does it run on an agarose gel? When you ignite a speck of it on a clean metal spatula - does it burn or does it just sit there (and what color does it become). Normally you can prepare DNA-protein complexes in a variety of ways, including direct addition, concentration, counterdialysis, etc. Regards, Artem -Original Message- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of E rajakumar Sent: Saturday, June 21, 2008 5:48 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] query on DNA-protein complex preparation for crystallization Dear All Sorry for non-crystallography question. I have synthesized two complementary strands of 16 bases in length for making duplex DNA and co-crystallization with DNA binding protein. I have mixed two complementary strands of 1:1 molar ratio (0.5 mM) in water and concentrated to 1.5 mM (Duplex), while concentrating solution becomes viscous and turned to white precipitate. However, adding 2 mM Magnesium chloride followed by annealing (heating at 90C for 10 minutes and followed by cooling to room temperature) did not help to dissolve the white precipitate. Please can you give me suggestions on following queries? 1.How do I dissolve white precipitate? Is increasing divalent cation or keeping duplex in particular pH could help in dissolving the precipitate? 2.How do I prepare DNA-protein complex? I mean, can I mix diluted DNA and protein in 1:1 molar ratio and concentrate further? Any guidance in this regard will be appreciated. Sorry, foregot to mention that any references in this regards will be great help. Thank you in Advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Send instant messages to your online friends http://uk.messenger.yahoo.com E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Send instant messages to your online friends http://uk.messenger.yahoo.com
Re: [ccp4bb] query on DNA-protein complex preparation for crystallization
Hi Thank you for the mail. It seems your correct. A(260 nm)/A(280) of one oligo is around 1.0 and peak is around 272. Other oligo's(260 nm)/A(280) is around 1.5. Can I know what is the absorbance peak of base protecting N-benzoyl group. Is it possible to do deprotection of base after mixing complementary strands? Can you suggest me what is the volume of ammonium hydroxide will be used for 1uM oligo of 16 bases in length and how much time heating shoul be done? thanking you rajakumara --- William Scott [EMAIL PROTECTED] wrote: Check the purity of the DNA in solution: A(260 nm)/A(280) = 1.8 for fully deprotected DNA, and you should see a nice clean simple curve with a peak very close to 260 nm. Check it on a denaturing gel. Smearing indicates incomplete deprotection. This is usually the cause of solubility problems. Sometimes resuspending in a strong cationic buffer (say 100 mM Tris pH 8.5) might be required. For crystallization it is probably best to have Na+ or K+ as a counterion, rather than Mg++. So you need to dialize against a high concentration of monovalent salt first, not just deionized water. On Jun 22, 2008, at 9:10 AM, E rajakumar wrote: Hi Artem Evdokimov Thank you for the mail. I have synthesized DMT-on oligos in our laboratory. Deprotection was performed treating with ammonium hydroxide for 15 hours at 55C. Then, DMT-on oligo was separated from off using RPHPLC. DMT was cleaved by treating with 20% glacial acetic acid for one hour. Then, DMT-off DNA was separated from DMT, again using RPHPLC. Lyophilized DMT-off oligos were dissolved in 3 mL of Milli Q water and dialysed against 2 L of milli Q water for 4hrs by changing water 2 times. Then complemntary oligos are concentrated around 1.0 mM and mixed them and concentrated further to 1.5 mM (duplex). 2 mM (final concentration) of Magensium chloride was added to oligos and concentrated to half of the volume. While concentrating oligos become viscos and white precipitate. however, annealing did not help to dissolve the white precipitate. I kept oligos in distilled water, without adusting pH. Please can you mail if I iginite DNA on metal spatual, eiether burns or not, what it indicates? Thanking you Rajakumara [EMAIL PROTECTED] wrote: Hi, How did you synthesize the DNA? I assume external vendor (so few people make their own these days)? How was the DNA purified? Sometimes if only a 'desalting' step is used there may be 'other chemicals' in the mix. Also, what pH was your DNA at, and in what buffer (if any)? If your DNA degraded you may have Pi in solution, which forms insoluble precipitates with many counterions. So, first of all I would check your white precipitate - does it dissolve in anything at all? If it does dissolve, what pH does it have? Does it run on an agarose gel? When you ignite a speck of it on a clean metal spatula - does it burn or does it just sit there (and what color does it become). Normally you can prepare DNA-protein complexes in a variety of ways, including direct addition, concentration, counterdialysis, etc. Regards, Artem -Original Message- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of E rajakumar Sent: Saturday, June 21, 2008 5:48 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] query on DNA-protein complex preparation for crystallization Dear All Sorry for non-crystallography question. I have synthesized two complementary strands of 16 bases in length for making duplex DNA and co-crystallization with DNA binding protein. I have mixed two complementary strands of 1:1 molar ratio (0.5 mM) in water and concentrated to 1.5 mM (Duplex), while concentrating solution becomes viscous and turned to white precipitate. However, adding 2 mM Magnesium chloride followed by annealing (heating at 90C for 10 minutes and followed by cooling to room temperature) did not help to dissolve the white precipitate. Please can you give me suggestions on following queries? 1.How do I dissolve white precipitate? Is increasing divalent cation or keeping duplex in particular pH could help in dissolving the precipitate? 2.How do I prepare DNA-protein complex? I mean, can I mix diluted DNA and protein in 1:1 molar ratio and concentrate further? Any guidance in this regard will be appreciated. Sorry, foregot to mention that any references in this regards will be great help. Thank you in Advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Send instant messages to your online friends http://uk.messenger.yahoo.com E. Rajakumara Postdoctoral Fellow
[ccp4bb]
Dear All Sorry for non-crystallography question. I have synthesized two complementary strands of 16 bases in length for making duplex DNA and co-crystallization with DNA binding protein. I have mixed two complementary strands of 1:1 molar ratio (0.5 mM) in water and concentrated to 1.5 mM (Duplex), while concentrating solution becomes viscous and turned to white precipitate. However, adding 2 mM Magnesium chloride followed by annealing (heating at 90C for 10 minutes and followed by cooling to room temperature) did not help to dissolve the white precipitate. Please can you give me suggestions on following queries? 1.How do I dissolve white precipitate? Is increasing divalent cation or keeping duplex in particular pH could help in dissolving the precipitate? 2.How do I prepare DNA-protein complex? I mean, can I mix diluted DNA and protein in 1:1 molar ratio and concentrate further? Any guidance in this regard will be appreciated. Thank you in Advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Send instant messages to your online friends http://uk.messenger.yahoo.com
[ccp4bb] query on DNA-protein complex preparation for crystallization
Dear All Sorry for non-crystallography question. I have synthesized two complementary strands of 16 bases in length for making duplex DNA and co-crystallization with DNA binding protein. I have mixed two complementary strands of 1:1 molar ratio (0.5 mM) in water and concentrated to 1.5 mM (Duplex), while concentrating solution becomes viscous and turned to white precipitate. However, adding 2 mM Magnesium chloride followed by annealing (heating at 90C for 10 minutes and followed by cooling to room temperature) did not help to dissolve the white precipitate. Please can you give me suggestions on following queries? 1.How do I dissolve white precipitate? Is increasing divalent cation or keeping duplex in particular pH could help in dissolving the precipitate? 2.How do I prepare DNA-protein complex? I mean, can I mix diluted DNA and protein in 1:1 molar ratio and concentrate further? Any guidance in this regard will be appreciated. Sorry, foregot to mention that any references in this regards will be great help. Thank you in Advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Send instant messages to your online friends http://uk.messenger.yahoo.com
[ccp4bb] 5-methy 2-deoxy Cytidine modified DNA duplex
Dear CCP4 users Sorry for non CCP4 topic. I want to know a company which can synthesize 5-methyl 2-deoxy Cytidine modified DNA duplex. Thank you in advance Rajakumara E. Rajakumara Postdoctoral Fellow Strcutural Biology Program Memorial Sloan-Kettering Cancer Center New York-10021 NY 001 212 639 7986 (Lab) 001 917 674 6266 (Mobile) Send instant messages to your online friends http://uk.messenger.yahoo.com
[ccp4bb] Purification using GST column at low pH
Dear All Sorry for non-crystallography question. I am purifying a protein using GST column. The gel filtration chromatography is indicating that the protein is aggregating at pH more than 7.0. I am planning to purify the protein at the pH around 6.0. So, I want to know the affinity of GST-fusion protein to the GST column and also activity of the Prescission Protease at pH 6.0. Thank you in advance Rajakumara Send instant messages to your online friends http://uk.messenger.yahoo.com
