Secondary Antibody: Goat anti Rabbit IgG Fc

The Biomat product is a 96 well coated microplate with goat anti  rabbit IgG  and a protein to block non-specific binding sites and to maintain stable activity.

Affinity purified goat anti rabbit IgG specifically binds the Fc region of  rabbit immunoglobulins , with minimal cross-reaction to human  serum proteins.

These plates are ideal for binding assays when available antibodies are in low quantities or they denature and become inactive upon direct adsorption  to polystyrene plates.

 

Features of goat anti rabbit IgG antibody coated plates:

 

Product specifications

Components

Individually pouched 96-well microplates, configured in 12 removable 8-well strips.

Coating

Affinity purified Goat anti rabbit IgG Fc  is coated using 100 µl/well. The strips are post-coated ( blocked ) for low non specific binding and long-term stability.

Binding capacity   

Microplate was saturated with rabbit IgG at a concentration of  1.0 µg/ml ( 100 ng/well )  in an ELISA format using goat anti rabbit IgG ( H+L )-HRP as detector and TMB as substrate ( see Figure 1 for data and experiment details ).

The Biomat Goat anti rabbit IgG Fc  microplate shows  a nominal binding capacity of ~  0.625 pmol /well of rabbit IgG 

Sensitivity  

Rabbit IgG was detected at a concentration significantly above background in an ELISA format using goat anti rabbit IgG ( H+L )-HRP as detector and TMB as substrate ( see Figure 1 for data and experiment details ).

The Biomat Goat anti rabbit IgG Fc  microplate shows  a sensitivity of  ~  0.01 µg/ml of  rabbit IgG.

Uniformity

Microplates show a CV% less than 5 when used as a catcher of  rabbit IgG in an ELISA format using goat anti rabbit IgG ( H+L )-HRPas detector and TMB as substrate.

Storage and Stability

The microplates, under the indicated storage conditions 2-8 °C, are stable until the expiration date printed on the label.
If opened, store in closed pouch with dessicant and use within the expiration date.

 

TECHNICAL NOTES N. 38 binding capacity and sensitivity test

1.     Add 100 µl of different concentrations of rabbit IgG ( from 0.025 to 4 µg/ml ) to the wells of goat anti rabbit IgG coated plate and incubate for 60 minutes at room temperature

2.     Empty the wells and wash with 0.1 M PBS pH 7.2,0.05% Tween® 20 four times

3.     Add 100 µl /well of Goat anti-rabbit IgG ( H+ L )-HRP ( Jackson ImmunoResearch code 111-035-003, diluted

1: 150.000 ) and incubate for 30 minutes at room temperature

4.     Empty the wells and wash with 0.1 M PBS pH 7.2,0.05% Tween® 20 four times

5.     Add 100 µl /well of TMB substrate solution and incubate 15 minutes at room temperature

6.     Stop the substrate reaction by adding 100 µl /well of sulphuric acid 1 N and read the optical density values at 450 nm

The data show that a plateau has got starting  with an IgG rabbit concentration of  1.0 µg/ml.

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

-          µg/well = 0.1 ( 100 ng/well )

-          pmol/well=  0.625 (this result is calculated considering the IgG M.W. = 160.000)

 

       The microplate sensitivity was calculated as the lowest rabbit IgG concentration higher than the mean optical 

       density plus  5 S.D. of  0 µg/ml  rabbit IgG concentration.

       Our experiment gave the following results :

-          0 µg/ml rabbit IgG optical density mean ( coming from 8 replicates ) = 0.121

-          standard deviation = 0.013

-          mean + 5 S.D. = 0.186

-          sensitivity = 0.010µg/well of rabbit IgG