Product Name: KAM-900P
Product Number: AR_KAM900P
Size: | 2 samples | | Price: | 869.00 |
| | | $US | |
Full Name: Kinex™ KAM-900P Antibody Microarray
Product Type Specific: Antibody microarray
Production Method: Validated antibodies are spotted with a microarray printer on to 3D matrix-coated glass slides in a staggered pattern duplicate spots distributed in 16 grids, twice in two fields to accommodate two separate samples. Median error range of ~12-14% for duplicates. Average of duplicate measurements of the same sample on different chips have a median error range of ~10%. Dynamic range of detection of up to 10,000-fold.
Storage Conditions: Upon receipt, the KAM-900P chip and dye-mix should be kept frozen at -20°C. Other components can be stored at 4°C for 1 month. For longer storage, the G25 spin-column and incubation chamber can be left at 4°C, but the other components should be frozen at -20°C.
Storage Stability: The KAM-900P Kit can be stored for at least 1 year if frozen at -20°C (G25 spin column and incubation chamber should not be frozen).
Array Use Description: The KAM-900P ANtibody Microarray Kit can be used to screen for changes in cell signalling protein phosphorylation (with ~613 phosphosite-specific antibodies) expression, and protein interactions (with ~265 pan-specific antibodies). In addition to discovering candidate proteins that are altered in experimental model systems, the Kit also allows for the identification of antibodies for biomarkers. The Kit permits users to perform screening of two cell or tissue lyates with duplicate measurements. The KAM-900P chip monitors the differential binding of dye- or biotin-labelled proteins in lysates prepared from cells and tissues including cells, fresh or frozen tissues and serum samples. The results can provide novel and useful insights into differences in protein expression, phosphorylation and protein-protein interactions, and define antibody reagents that can be used to follow up on these findings in immunoblotting, immunoprecipitation, ELISA and immunohistochemistry studies.
Array Probe Description: The KAM-900P chip uses ~878 distinct antibodies (~265 pan-specific and ~613 phosphosite-specific) for protein kinases and other cell signalling protein. These antibodies were tested in-house at KInexus and cherry-picked from over 5000 sourced from over 25 vendors with about half of the antibodies developed by and available directly from Kinexus.
Array Slide Type: Nexterion-P 3D matrix coated glass slides from Schott.
Array Slide Methodology: Single dye, non-competitive methodology - Two samples are labelled with the same dye and incubated separately on the same slide to eliminate differential protein-dye binding problems. The optional use of proprietary chemical cleavage of sample proteins lowers the risk of false positives from protein-protein interactions, loss of protein phosphorylation, and reduces differences in signal intensities due to differences in the size of target proteins.
Scientific Background: High content antibody microarrays are powerful tools that permit the mass profiling of hundreds of diverse proteins for alterations in protein levels, post-translational modifications and protein-protein interactions. Kinexus offers a chemical cleavage step that reduces the occurence of protein-protein interactions. With their high sensitivities, as little 25 µg amounts of cell or tissue lysate proteins are required, although 100 µg of lysate protein is recommended. With the KAM-900P chip, highly validated antibodies are printed on to the surface of a glass slide. Lysates proteins are either directly pre-labeled with a proprietary mixture of fluorescent dyes (or biotin), and subsequently captured on the microarray (If the lysate proteins are biotinylated, then the captured proteins are detected with fluorescent dye-labelled anti-biotin antibody). The intensity of the fluorescent signal at each antibody spot is a measure of how much protein was bound. Due to the possibilities of antibody cross-reactivities and protein-protein interactions in diverse cell/tissue types, the leads should be followed by Western blotting. Kinexus offers a parallel multi-immunoblotting service to confirm the results of our KAM antibody microarrays. Over 5000 antibody microarray analyses performed to date. Open access in KiNET-AM (www.kinet-am.ca) to full experimental data from over 2000 analyses available online for comparisons. Over 250 scientific peer-reviewed papers published using our antibody microarrays.