b'iBind western detection systemsTwo devices, one simple technology for western blot processingThe Invitrogen iBind western blot systems are simple, unpowered devices that automate immunodetection steps. The traditional manual process involves preparing and replacing multiple antibody and wash solutions over several hours in a tray containing the blot of interest. In contrast, iBind western systems allow all solutions to be prepared and loaded in the device at the start of the procedure, with subsequent steps proceeding automatically and uninterrupted by sequential lateral flow technology (SLF), i.e., simple capillary actionno electricity or batteries are required.The original iBind Western Device accommodates processing one mini blot at a time, and the iBind Flex Western Device accommodates processing one midi blot, two mini blots, or up to six vertically cut strips at a time for additional flexibility and throughput.Highlights:Antibody savingsuse up to 80% less primary antibody than with traditional tray-based format (Figure 5)Reproducibilityautomated processing provides improved blot-to-blot consistencyLoad and goprocess solutions using SLF technology, with no batteries, shakers, trays, or timers requiredGST-tagged proteins (two-fold dilution series) Figure 5. Comparison of western blots processed manually vs. IKK DDR2 FLT1 HCK with the iBind Flex Western Device. Samples containing GST-tagged 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 recombinant proteins were separated on Invitrogen NuPAGE 412%, 20-well midi gels in MOPS SDS running buffer and then transferred to iBind Flex nitrocellulose membranes using the iBlot 2 Gel Transfer Device. Blots Westernwere probed with identical concentrations of the same pair of primary System and secondary antibodies. The primary antibody was rabbit anti-GST diluted 1:500 (8 L in 4 mL iBind Flex Solution for the iBind system, 40 L in 20 mL for manual tray incubation). The secondary antibody was goat antirabbit IgG HRP diluted 1:600 (6.7 L in 4 mL iBind Flex Solution for the iBind system, 33.3 L in 20 mL for manual tray incubation). For Manual (tray) final detection, blots were incubated for 5 minutes in Thermo Scientific processing SuperSignal West Dura Extended Duration Substrate for visualization with an imaging system.Lanes 15: IKK (80 ng, 40 ng, 20 ng, 10 ng, 5 ng)Lanes 610: DDR2 (120 ng, 60 ng, 30 ng, 15 ng, 7.5 ng)Lanes 1115: FLT1 (40 ng, 20 ng, 10 ng, 5 ng, 2.5 ng)Lanes 1620: HCK (360 ng, 180 ng, 90 ng, 45 ng, 22.5 ng)View the how-to video:Learn more at thermofisher.com/ibind66 Western blotting thermofisher.com/proteinbiology'