Plates for Elisa and Research

Medium Binding

A hydrophobic surface suitable for passive adsorption of proteins with different grades of hydrophobicity.

 

FEATURES

Assays in which the adsorbed molecule present large hydrophobic regions, such as antibodies.

Furthermore this surface is highly selective and shows high affinity towards hydrophobic polypeptides that present a molecular weight > 10kDa.

 

PERFORMANCES

A coating of antibodies of IgG class was carried out to evaluate its usefulness along with to evaluate the binding capacity of this surface.

 

Method 15

 

Method 15 is an indirect method with human IgG coated on medium binding plates and then revealed through an AHIgG/POD conjugate.

 

1.     dispense 100 ml/well of different concentrations of human IgG diluted in 0.1M PBS pH 7.2 : 1-2-3-4-5 µg/ml and incubate overnight at         4°C

2.     wash 3 times with 0.1M PBS pH 7.2 + 0.05%Tweenâ 20

3.     dispense 150 ml/well of BSA 1% in 0.1M PBS pH 7.2  and incubate 2 hours at R.T. for blocking the remaining active sites

4.     wash 3 times with 0.1M PBS pH 7.2+ 0.05% Tweenâ 20

5.     dispense 100 ml/well of Goat Anti-HIgG-HRP conjugate and incubate 30’ at R.T.

6.     wash 3 times with 0.1M PBS pH 7.2 + 0.05% Tweenâ 20

7.     dispense 100 ml/well of TMB

8.     after 30' stop the reaction with H2SO4 1 N

9.     reading is made at 450 nm

                                                                        MEDIUM BINDING CAPACITY.pdf

The data  show that a plateau has got starting  with an IgG concentration of 2.0 µg/ml.

This concentration means the well binding capacity we can express as:

-          µg/well = 0.200 ( 200 ng/well/100µl)

As 100 µl of liquid, in term of area , represent 1 cm2 it is possible to state that the binding capacity is close to 200 ng/ cm2. These data are well correlated with other experiments carried out with an unmodified polystyrene surface.

96 well microplates

Material: Polystyrene
Quantity per pack: 25 - Quantity per case: 200

1x8 flat bottom wells strips on 12 x 8 frame

1x8 flat bottom wells breakable strips on single well holding frame

96 wells plate - flat bottom