Luminex xMAP Antibody Coupling Kit Manuel d'utilisateur télécharger pdf (Page 19)

Page 15 of 20

4. Prepare a solution of phycoerythrin-labeled anti

-species IgG detection antibody at 4

μg/mL in PBS-1% BSA. Prepare a 1:2 dilution series of that detection antibody

solution to a concentration of 0.0625 μg/mL as shown in the following table.

NOTE: F

or optimal results use Costar round-bottom 96-well plates and Luminex

Magnetic Plate Separator with MagPlex microspheres.

5. Aliquot 50 μL of

the microsphere solution prepared in Step 3 into two entire columns

of wells of the plate (duplicate sets of 8 wells, 16 wells total).

6. Add 50 μL of

PBS-1% BSA, as a blank sample, into the wells in Row A containing

the microsphere solution.

7. Add 50 μL of each of the di

luted detection antibody solutions prepared in Step 4 into

the appropriate wells of the plate (as shown in the plate layout below).

1 2 3 4 5 6 7 8 9 10 11 12

Blank Blank

1:64 1:64

1:32 1:32

1:16 1:16

1:8 1:8

1:4 1:4

1:2 1:2

1:1 1:1

Example plate layout using columns 1 & 2

Dilution Tube

Volume of

PBS-1% BSA

Volume of Detection

Antibody

Concentration

1:1 - - 4 μg/mL

1:2 500 μL 500 μL from Tube 1:1 2 μg/mL

1:4 500 μL 500 μL from Tube 1:2 1 μg/mL

1:8 500 μL 500 μL from Tube 1:4 0.5 μg/mL

1:16 500 μL 500 μL from Tube 1:8 0.25 μg/mL

1:32 500 μL 500 μL from Tube 1:16 0.125 μg/mL

1:64 500 μL 500 μL from Tube 1:32 0.0625 μg/mL

1 2 ... 14 15 16 17 18 19 20 21 22 23 24

Pas de commentaire

Luminex xMAP Antibody Coupling Kit Manuel d'utilisateur Page 19