microarrays showing binding of the six ankyrin domains from TRPV1 to Orai1 carboxyl terminus

8 microarray spots (replica) illustrating the binding of the ankyrin domains 1-6 from TRPV1 (amino acid sequences shown below) to the carboxyl terminus from Orai1. Each microarray raw file in tif format shows 8 repeats (microarray spots) for each ankyrin peptide.Orai1-COOH-dsRED was deposited on each microarray spot. Attachment of each peptide from Orai1 to the microarray was monitored by measuring dsRED fluorescence (shown in red).Files are in RGB TIF format and named accordingly to the ankyrin domain explored (1-6).
Each RGB tif file contains 3 channels: Blue (empty), Red (signal from Orai1-COOH-dsRED) and Green (signal from the binding of the ankyrin peptides fused to GFP). Red channel used to monitor the amount of Orai1-COOH-dsRED deposited on each microarray spot to ensure even distribution.
Ankyrin domain sequences from human TRPV1 are:
ANK1 110..152 (42 aa) RLYDRRSIFDAVAQSNCQELESLLPFLQRSKKRLTDSEFKDPE

ANK2 153..199 (46 aa) TGKTCLLKAMLNLHNGQNDTIALLLDVARKTDSLKQFVNASYTDSYY

ANK3 200..246 (46 aa) KGQTALHIAIERRNMTLVTLLVENGADVQAAANGDFFKKTKGRPGFY

ANK4 247..282 (35 aa) FGELPLSLAACTNQLAIVKFLLQNSWQPADISARDS

ANK5 283..331 (48 aa) VGNTVLHALVEVADNTVDNTKFVTSMYNEILILGAKLHPTLKLEEITNR

ANK6 332..358 (26 aa) KGLTPLALAASSGKIGVLAYILQREIH
All six peptides fused to the Green Fluorescent Protein (GFP) used to detect the binding of the ankyrin peptides to the Orai1-COOH-dsRED deposited on the microarray spots.Peptide binding detected by Total Internal Reflection Fluorescence Microscopy (TRIFM) using a closed chamber and a TIRFM system developed by TIRFLabs. Dual color emission filters for green (GFP) and red (dsRED) were switched to acquire the data and stored in RGB tif format.For details on the protocols please refer to:

A platform for combined DNA and protein microarrays based on total internal reflection fluorescence.
Asanov A, Zepeda A, Vaca L.
Sensors (Basel). 2012;12(2):1800-15. doi: 10.3390/s120201800. Epub 2012 Feb 9.
PMID: 22438738