Association of Biomolecular Resource

Ex situ (also known as spotted or printed) arrays have become very popular formats, especially for the building of custom noncommercial arrays used primarily by academic laboratories [see Association of Biomolecular Resource Facilities (ABRF) surveys on microarrays atwww.abrf.org]. The printed cDNA microarray was largely developed from gene expression work originating in the laboratories of RO. Brown and R.W. Davis at Stanford University (Schena et al., 1995). Plans for the construction of the microarrayer and split pin designs were available at the Brown lab website at http //cmgm.stanford.edu/ pbrown/mguide/index.html. This enabled researchers to prepare their own microarrays appropriate for their particular experiments. 

The Association of Biomolecular Resource Facilities (ABRF) has been addressing the identification and ultimate resolution of such difficulties by collaborative trials [1-7]. The purpose of the 1994 trial was to discriminate between hydrolysis and the chromatographic analysis as a possible major source of errors, and additionally to test cystine analyses as part of a continuing effort. 

The Association of Biomolecular Resource Facilities (ABRP) Protein Sequence Research Committee was established in 1988 in order to provide individual laboratories with a means of self-evaluation. Each year test samples have been distributed, enabling laboratories an opportunity to monitor their performance in areas such as sample handling, insuument operation/optimization, and data interpretation. In previous years these samples have focused on sensitivity of protein sequencing (1, 6), sample heterogeneity (2, 8), protein-bound peptides on PVDF membrane or in solution (3, 4), post-translational modifications (5), identification of cysteine and tryptophan (7), and length of sequence assignment (8). 

Abbreviations used AAA, amino acid analysis ABRF, Association of Biomolecular Resource Facilities AQC, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate FMOC, N-(9-fluorenylmethoxycarbonyl) OPA, o-phthaldialdehyde PITC, phenylisothiocyanate tpis, triosephosphate isomerase. 

The Peptide Synthesis Research Committee of the Association of Biomolecular Resource Facilities (ABRF) conducts anonymous studies to evaluate the ability of ABRF member laboratories to s ynthesize and characterize test peptides (1-5). The committee has also conducted studies which provided an opportunity for our member laboratories to attempt new synthetic methods and evaluate new analytical technologies. Previous studies by this committee have shown that peptide assembly and cleavage are no longer significant problems in most core laboratories. Therefore, for its 1996 study, the ABRF Peptide Synthesis Research Committee sought to assess the extent to which racemization occurs during peptide assembly in peptides synthesized by our member resource laboratories. 

MW Crankshaw, GR Grant. Identification of modified PTH-amino acids in protein sequence analysis. Presented at the Association of Biomolecular Resource Facilities, 1993. 

Figure 6.7 Schematics of single-quadrupole (a) and quadrupole ion trap (b) mass analyzers. (Reproduced from Jonscher and Yates with permission from ABRF News the Association of Biomolecular Resource Facilities copyright 1996.)...

Tarr, G.E. Paxton, R.J. Pan, Y.C.E. Ericsson, L.H. Crabb, J.W. Amino acid analysis 1990 The third collaborative study from the Association of Biomolecular Resource Facilities (ABRF). In Techniques in Protein Chemistry, 2nd Ed. Villafranca, J.J., Ed. Academic Press San Diego, CA, 1991, 139-150. 

Laboratones not set up for structural characterization of proteins can find assistance for ammo acid analysis, Edman degradation and mass spectrometry in the Yellow Pages of the Membership Directoiy of the Association of Biomolecular Resource Facilities (ABRF, 9650 Rockville Pike, Bethesda, MD)... 

FIGURE 18.8 A schematic of Dole s original electrospray nozzle. (Reproduced from Fenn, J.B., J. Biomol. Tech., 13, 101, 2002. With permission of the Association of Biomolecular Resource Facilities.)... 

FIGURE 18.12 ESI-MS spectra for medium molecular weight peptides but well beyond the nominal capabiUty of a quadrupole mass analyzer if z = 1. This is made possible by the careful design of the electrospray nozzle and the attached H ions which brings the im/z ) ration into range of the quadrupole mass analyzer. (Reproduced from Perm, J.B., J. Biomol. Tech., 13, 101, 2002. With permission of the Association of Biomolecular Resource Facilities.)... 

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