The yeast proteome collection was derived from the yeast clone collection of 5800 yeast ORFs. The identity of each clone was verified using 5' end sequencing. Expression of glutathione-S-transferase (GST)-tagged protein by each clone was tested using Western blotting and detection with an anti-GST antibody. The 4088 clones that passed both quality control measures were purified using high-throughput affinity chromatography as previously described. A contact-type printer (Omnigrid, Genomic Solutions) equipped with 48 matched quill-type pins was used to deposit each of 4,088 purified yeast proteins along with a set of control elements in duplicate spots on 1" x 3" glass slides. The printing of these arrays was carried out in a cold room under dust-free conditions in order to preserve the integrity of both samples and printed microarrays. Before releasing these protein microarrays for use, each lot of slides is subjected to a rigorous quality control procedure, including a gross visual inspection of all the printed slides for imperfections. The second step consists of a more detailed characterization of each spot on the array. Since each of the proteins is tagged with GST, this QC procedure is accomplished by using an antibody detection protocol specific for GST. This procedure measures the variability in spot morphology, the number of missing spots, the presence of control spots, and the amount of protein deposited in each spot.
Catalog number
PA0121011
Support
glass
Coating
nitrocellulose
Description
The Yeast ProtoArray™ high-density functional protein microarray is the first microarray product that enables rapid elucidation of proteinprotein interactions on a proteome scale. The Yeast ProtoArray™ contains 4,088 S. cerevisiae open reading frames (ORFs) expressed as 5'-GST fusions, purified and spotted in duplicate on a 1 inch x 3 inch nitrocellulose-coated slide (Figure 1).With the Yeast ProtoArray™ PPI Kit, you can identify novel protein interactions in a single day, instead of the several weeks required using alternatives such as yeast two-hybrid systems. In addition, protein interactions can be easily assayed under different conditions to better understand interaction dynamics.
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