What's New?
Spring Break Closure for PCL
The PCL will be closed March 16-17th for spring break. Samples that need to be concluded before that time should arrive by March 7th.
Posted: Wednesday, February 22, 2006, 02:48PM EST
Hydroxyproline and Hydroxylysine Assay
Hydroxyproline and hydroxylysine, found primarily in connective tissues, can be quantitated by amino acid analysis in a single run with tandem detection by UV and flourimetry. Both the cis- and trans- forms of OH-Pro, being secondary amino acids, are derivatized by FMOC in our system and are best monitored at Ex=266/Em=324 for specificity. OH-Lys, being a primary amino acid, is derivatized by OPA (and elutes as two peaks, the singly and doubly derivitized molecule) and is best monitored at 338 nm for sensitivity.
Posted: Wednesday, May 19, 2004, 04:13PM EDT
Phosphoserine Assay
Although no specific Serines can be identified as being phosphorylated, a partial hydrolysis at 110* C for 2 hours allows us to measure the ratio of p-Ser to Ser in peptides and proteins. Using phosphorylated Neurogranin as a standard, the amount of p-Ser at time=zero can be calculated for a protein within a 5% RSD of within-assay variation. The assay requires the use of flourometric detection at Ex=340, Em=450 for specificity.
Posted: Monday, May 10, 2004, 12:28PM EDT
PCL has new Faculty Director
Dr. Donald Pettigrew, Professor of Biochemistry and Biophysics, is the new Faculty Director of the PCL as of October 1, 2003. Dr. Pettigrew will be taking over the Faculty Directorship of the laboratory from Dr. Marty Scholtz who has held the position for the last several years but must step down to accommodate his new responsibilities as Acting Head of Medical Biochemistry and Genetics in the Texas A&M Health Sciences Center. We welcome Dr. Pettigrew on board and look forward to a productive relationship.
Posted: Monday, January 12, 2004, 04:30PM EST
PCL Leads Proteomics Workshop in Mexico City
Larry Dangott and the staff of the PCL have been invited to present a workshop on 2D gel electrophoresis and proteomics at the Centro de Investigacion y de Estudios Avanzados (CINVESTAV) in Mexico City from November 10-14, 2003. This workshop will provide hands-on training and access to state-of-the-art techniques and instrumentation available at Texas A&M, nurturing a growing relationship between Texas A&M University and one of Mexico’s premier research institutes. This workshop will help to develop new research collaborations and attract new students and postdoctoral fellows from our neighbor to the South, as well as expanding research opportunities and international experiences for students at Texas A&M University. Interactions with CINVESTAV grew from efforts of the Department of Biochemistry & Biophysics and the College of Agriculture and Life Sciences, led by Drs. Jorge Cruz-Reyes and Sumana Datta, and CINVESTAV faculty member, Prof. Gabriel Guarneros Peña, to strengthen the basic and applied research ties between our two institutions. The PCL is supported by the Office of the Vice-President for Research, the Center for Environmental and Rural Health, and the Center for Advanced Biomolecular Research and provides support for research in protein chemistry, proteomics, and molecular biology at Texas A&M.
Posted: Monday, January 12, 2004, 04:29PM EST
New Glucosamine Assay
Jinny Johnson of the PCL has implemented a new, rapid, quantitative assay for the glucosamine using reverse phase HPLC. Upon request from Dr. Potter from the Department of Animal Science at TAMU, Jinny has developed and validated a method to determine levels of the amino-sugar, glucosamine, in equine serum. The method is relatively robust and sensitive down to several micrograms. This representative chromatogram shows how the method can separate glucosamine anomers from other amino acids. The method is quantitative. Please call (979) 845-2433 or e-mail jinny@tamu.edu for details.
Posted: Monday, January 12, 2004, 04:27PM EST
New Tryptophan Assay
Jinny has developed a more reliable assay for Tryptophan based on literature references. This procedure is offered as a separate analysis from standard amino acid analysis as it requires a larger amount of sample. You will need to submit sample for our standard assay and our Tryptophan assay if you wish to analyze all amino acids. Please contact Jinny (jinny@tamu.edu) or call the PCL for additional details.
Posted: Monday, January 12, 2004, 04:24PM EST
New O'Farrell Gel System Uses SDS to Solubili
Breaking all dogma, we have been applying samples dissolved in SDS to O’Farrell style gels AND GETTING EXCELLENT RESULTS. Following the lead of the Kendrick Labs in Wisconsin, we are using SDS sample buffer (Laemmli) to solubilize hard-to-solubilize samples and applying them to tube gel IEF procedures. From this we get a fine pattern of spots from samples that ordinarily give few or none when treated with urea and subjected to IPGDryStrip analysis. Consider this as an alternative for your difficult samples. There are some volume and concentration restrictions that apply to this method, so please contact Larry (ljdangott@tamu.edu) or call the lab for additional details. See Electrophoresis FAQ for examples of these gels.
Posted: Monday, January 12, 2004, 04:23PM EST
New Pierce FeBABE Articifial Protease for Map
This product is useful in mapping Protein-Protein interactions and a new addition to the ProFound product line of ‘pull-down’ kits used to identify protein-protein interactions. Taking advantage of work by Claude Meares (University of California, Davis), Pierce has created a new kit that uses the iron-chelating agent FeBABE that cleaves proteins at sulfhydryl groups on cysteine residues. In this system, a ‘bait’ protein is labeled with FeBABE and the ‘prey’ is end-labeled with hemagluttinin. After the two proteins are allowed to interact, chemical cleavage is induced and the fragments analyzed by Western Blot analysis to profile the pieces. Visit Pierce Chemical Co. (http://www.piercenet.com) to learn more about it. Go to the product now.
The PCL is working with MIT scientists to develop living, artificial cartilage. See how Tissue Engineering is striving to develop replacement tissues for reconstruction of human body parts. Go to the page now.
Posted: Monday, January 12, 2004, 04:20PM EST
Improved In-Gel Digestion Capabilities
The PCL has added a new system for improving and accelerating in-gel digestion of proteins. The Montage In-Gel DigestZP Kit produced by Millipore is a fast, convenient and reproducible method to digest proteins and purify peptides in up to 96 polyacrylamide gel pieces simultaneously for MS analysis. The procedure has been optimized for polyacrylamide stained gels of 1-1.5 mm in thickness and can be adapted for silver stained or Coomassie stained gels. Proteins may also be reduced and alkylated in the procedure.
We are currently using the technique to identify protein spots from 2D gel separations of seminal plasma proteins from stallion in collaboration with Dr. Dickson Varner of the TAMU Vet School and Drs. David Russell and William Russell in the Department of Chemistry at TAMU. We are also using the Montage to study stallion sperm plasma membrane proteins with Drs. Larry Johnson and Dickson Varner of the TAMU Vet School. The system was purchased with funds provided to Dr. Johnson by the Texas Equine Research Account.
Visit us or the Millipore Website to learn more about this procedure.
Posted: Monday, January 12, 2004, 04:19PM EST
New Assays
We have expanded the list of amino acids that we are able to analyze and quantitate. Jinny Johnson (Amino Acid Analysis Specialist) has validated assays for cis- & trans-Hydroxyproline (found in many connective tissues), desmosine (a crosslinked form of lysine) and Diaminopemilic acid (DAPA), an amino acid not found in proteins but commonly found in the proteoglycan layer of many bacteria. We will be posting additional information (and representative data) concerning these assays in the near future
Posted: Monday, January 12, 2004, 04:17PM EST
New 2D Gel System
2D gel electrophoresis requests are increasing at a high rate. Additionally, the types of samples being analyzed is diversifying. We are currently working on projects from such diversified systems as Arabidopsis and bacterial proteins. One project with Dr. Ron Martens (Department of Large Animal Science) is to identify cell-surface antigens on Gram-positive bacteria (Rhodococcus equi) that are involved in specific foal diseases. We are also using proteomics approaches in collaboration with the Laboratory for Biological Mass Spectrometry (TAMU, Chemistry) to identify soluble proteins in stallion seminal plasma with Dr. Dickson Varner (Dept of Large Animal Surgery). In another collaboration with Dr. Jim Hu (Department of Biochemistry & Biophysics), we are using rpHPLC and robotics to improve high-throughput MALDI-TOF analysis of E. coli proteins from whole cell lysates grown under different growth conditions.
DryStrip™ Immobilized pH Gradient technology is a very popular technique for first dimension Isoelectric focusing and is commonly used in our laboratory. Often, however, the technique is inappropriate and new approaches and different techniques are required to obtain the best results for a given sample. In response, we are offering TUBE GEL Isoelectric Focusing using traditional carrier Ampholytes for certain samples. Often this technique is better suited to difficult sub-groups of proteins (membrane proteins). Be sure and call us to discuss your particular needs.
Posted: Monday, January 12, 2004, 04:13PM EST
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