Multiscreen Filter Plates from EMD Millipore

April 09, 2013

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Stanford University
Structural Biology
Post-Doc

Image showing the Multiscreen filter plate mounted on the vacuum manifold of the Qiagen 9600 BioRobot.

Company:

EMD Millipore

Product Name:

Multiscreen Filter Plates

Catalog Number:

MSDVN6510
Image

Multiscreen filter are non-sterile, 0.65uM hydrophilic, low protein binding plates with 96 wells. Ideal for use in multiplex assays, these plates allow the user to standardize washing procedures across several wells using a multi-channel pipette. In our lab we use these plates in an assay that measures binding of recombinant Killer-cell Immunoglobulin-like Receptor (KIR) proteins against a panel of 96 HLA class I allotypes annealed to individually identifiable beads. The filter plates are used in the washing and incubation steps of this assay, retaining the beads whilst allowing the wash solution to pass through the filter. The plates are used with a vacuum source, in our case a Qiagen Bio Robot 9600 that has a vacuum manifold.

Experimental Design and Results Summary

Application

Multiplex bead assays. Multiscreen 96 well filter plates are used for the incubation and washing of recombinant KIR proteins and secondary antibodies bound to single antigen HLA class I beads.

Starting Material

Recombinant KIR protein, PBS, wash buffer, 96 well Multiscreen filter plate, Vacuum manifold (e.g Qiagen BioRobot 9600).

Protocol Overview

For each recombinant protein to be tested, use one well of the 96-well plate; 300ul of PBS should be added and aspirated using the vacuum. This acts to pre-wet the filter prior to addition of the protein to be tested. Add 50 ul (100ug/ml) of each protein to each pre-wet well of the 96-well plate. Add 3ul single antigen HLA class I beads (e.g One Lambda Labscreen, # LS1A04) to each well. Incubate, covered and shaking gently for 60min at 4°C. Following incubation, add 200ul wash buffer to each well and gently pipette up and down to resuspend and wash the single antigen beads. Aspirate the wash buffer using the vacuum manifold. Repeat this step four times. Add an appropriate secondary antibody (e.g. anti-human IgG1 Fc – One Lambda, #LS-AB2) in 100ul PBS to each well following the last aspiration. Incubate for 60min, covered and shaking at 4°C. Repeat the wash steps and resuspend the beads in each well with 100ul PBS. The beads are then ready to be analyzed on the Luminex platform.

Tips

Be careful not to puncture the delicate filter membranes with the pipette tips – this most commonly occurs during the wash steps. We use a multi-channel pipette for the wash steps to ensure that each well receives an identical washing regimen. The pressure of the vacuum should not exceed 100mmHg and the filter plates should not run dry during this process as this likely damages the single antigen beads. Samples should be transferred to a v-bottom 96-well plate for analysis as the Multiscreen filter plates do not have sufficient depth to be compatible with a Luminex 100 reader.

Results Summary

These filter plates are ideal for the multiplex assay described above. The 96-well format lends itself to a multiplex assay format in which it is important that each well receives as near to identical wash procedure as possible. These plates are robust enough to be used for both incubation and washing.

Additional Notes

None.

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Image Gallery

Image showing the Multiscreen filter plate mounted on the vacuum manifold of the Qiagen 9600 BioRobot.

Summary

The Good

Robust plate ideal for multiplex bead based assays.

The Bad

Filters can be ruptured by pipette tips leading to loss of sample during washing and sample transfer.

The Bottom Line

A useful adjunct for multiplex assays that saves considerable time by eliminating multiple centrifuge steps.

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