Luminex Flock Monitor User Manual

Flock Monitor

IBDV · NDV · IBV · REO

Quick Guide

This Quick Guide is not intended to replace the xMAP® Flock Monitor (IBDV, NDV, IBV, REO) Assay Package Insert (document number 89-6000000-070 rev. 9). Please refer to the Package Insert for complete background, instructions, warnings and procedure precautions.

Sample, Reagent and Plate Preparation

Before setting up the Flock Monitor assay plate, use the following procedures to prepare fresh reagents and sample dilutions. Allow all components to equilibrate to room temperature, and prepare aliquots and dilutions immediately prior to performing the assay.

SAMPLES

CONTROLS

MICROSPHERE

MIX

SA-PE

CONJUGATE

DETECTION

ANTIBODY

• Dilute all samples 1:500 using Sample DILUENT. You will need 50 µL of each diluted

sample per well.

• IMPORTANT - Vortex samples before use and after dilution.

• Dilute CONTROLS labeled (-) and (+) 1:500 using Sample DILUENT. You will need 50 µL

of each diluted control per well.

• IMPORTANT - Vortex controls before use and after dilution.

• Calculate the volume of microspheres you will need with the following formula: Required microsphere volume = (0.05 mL) x (number of wells) + (10%)

• Aliquot the required microsphere volume into another tube or reagent trough, shield

from light, and set aside for use in assay setup.

• Calculate how much conjugate you will need with the following formula: Required SA-PE conjugate volume = (0.10 mL) x (number of wells*) + (10%)

• Aliquot this amount into a reagent trough, then cover with foil to protect from light.

• Promptly return SA-PE conjugate to 2-8°C storage.

• Calculate how much conjugate you will need with the following formula: Required conjugate volume = (0.10 mL) x (number of wells*) + (10%)

• Aliquot this amount into a reagent trough, then cover with foil to protect from light.

• Promptly return detection antibody to 2-8°C storage.

WASH

BUFFER

MAGNETIC

SEPERATION

DEVICE

*Consult the product insert for more information

• WASH Buffer is provided ready-to-use in nal dilution.

• May be a LifeSep 96F hand held magnet for maual washing (see http://www.luminexcorp.

com/support/Magnetic%20Microspheres/Washing_Method_whitepaper.pdf for more details)

• May be a Bio Tek ELx405 or Tecan Hydroex washer equipped with a washer manufacturer provided magnet for automated washing

89-30000-00-261 Revision A - Page 1 of 2

Before You Start

1234 56789 10 11 12

Prepare reagents, samples and controls as described on page 1 of this Quick Guide. All steps below are performed

at room temperature, taking care to protect MICROSPHERES, SA-PE CONJUGATE, and DETECTION ANTIBODY

from light by wrapping in foil during incubation.

Allow liquid components to equilibrate to room

temperature for 30 to 60 minutes*

Warm up and calibrate Luminex instrument*

Vortex working MICROSPHERES 5 seconds

Sonicate 5 seconds

Add 50 μl MICROSPHERES to each well

SET UP

Vortex diluted CONTROLS and diluted SAMPLES

Add 50 μl CONTROLS or SAMPLE to

corresponding wells

Cover plate with opaque lid or foil

Shake plate at 800 rpm for 60 minutes

INCUBATEINCUBATE

WASH STEP - May be done with manual or automatic washer technique

Allow plate to sit on magnet for 60 seconds

Remove liquid from wells

WASH

Add 100 μl WASH to each well

Repeat WASH STEP one more time (total of two

washes)

Remove Final WASH from Wells

Plate Layout

Use this template for setting up your assay plate and

software batch on the Luminex 200 / FlexMAP 3D System.

Shake plate at 800 rpm for 30 minutes

INCUBATE

WASH STEP - May be done with manual or automatic

washer technique

Allow plate to sit on magnet for 60 seconds

Remove liquid from wells

WASH

Add 100 μl WASH to each well

Repeat WASH STEP one more time (total of two

washes)

Resuspend MICROSPHERES in 100μl WASH with

a pipette or plate shaker

Read assay results on a Luminex analyzer

READ

Add100 μl DETECTION ANTIBODY to each well

Cover plate with opaque lid or foil

DETECTION ANTIBODY

Shake plate at 800 rpm for 30 minutes

WASH STEP - May be done with manual or

automatic washer technique

Allow plate to sit on magnet for 60 seconds

Remove liquid from wells

WASH

Add 100 μl WASH to each well

Repeat WASH STEP one more time (total of two

washes)

89-30000-00-261 Revision A - Page 2 of 2

UNITED STATES

Luminex Corporation

Austin, Texas

Tel: +1.512.219.8020

Fax: +1.512.219.5195

Remove nal WASH from wells

Add 100 μl SA-PE CONJUGATE to each well

Cover plate with opaque lid or foil

CONJUGATE

*Consult the product insert for more information

CANADA

Luminex Molecular Diagnostics

Toronto, Ontario

Tel: +1.416.593.4323

Fa x: + 1.4 16. 593 .10 66

EUROPE

Luminex B.V.

Oosterhout,The Netherlands

Tel: +31.162.408333

Fax: +31.162.408337

www.luminexcorp.com

+ +

Protect microspheres, conjugate, and detection antibody

from prolonged light exposure throughout entire procedure.

CHINA

Luminex Shanghai Trading Co..

Shanghai, China

Tel: +21.616.50809

Fax:+21.616.50811

info@luminexcorp.com

USA: 1-877-8-Biovet (824-6838)

Canada & Intl: 1-888-8-Biovet (824-6838)

JAPAN

Luminex Japan Corporation, Ltd.

Tokyo, Japan

Tel: +81.3.5545.7440

Fax:+81.3.5545.0451

AUSTRALIA

BSD Robotics, A Luminex Company

Brisbane, Queensland

Tel: +61.7.3273.0273

Fax:+61.7.3273.0274