Coating Antigen Concentration (/100 µL/well)

Wave-length

(nm)

Conjugated Antibody Dilution

P/N of the Optical Density (OD) Reading at Different Levels of the Positive Serum Dilution

SEM

Blocked Wells

Non-blocked Wells

1:50

1:100

1:200

1:500

1:50

1:100

1:200

1:500

20 µg

580

1:2500

2.41^b1

2.43^a1

2.24^b2

2.32^b1,2

0.52⁴

0.58⁴

0.56^c4

1.38^c3

0.036

1:5000

2.55^b1

2.45^a2

2.45^a2

2.34^b3

0.55^5,6

0.57⁵

0.51^c6

1.49^a4

0.017

1:10,000

2.75^a1

2.53^a2

2.49^a2

2.47^a2

0.62⁵

0.60⁵

1.65^a3

1.53^a4

0.020

600

1:2500

1.85^d1

1.79^c1,2

1.67^e2

1.81^e1

0.58⁵

0.58⁵

0.76^b4

1.40^bc3

0.039

1:5000

1.99^d1

1.92^bc1,2

1.89^d1,2

1.87^cde2

0.55⁵

0.56⁵

0.76^b4

1.48^ab3

0.039

1:10,000

2.22^c1

2.02^b2

2.13^c1,2

1.97^c2

0.61⁴

0.59⁴

1.68^a3

1.52^a3

0.050

630

1:2500

1.83^d1

1.80^c1

1.58^e2

1.79^e1

0.55⁵

0.58⁵

0.72^b4

1.39^c3

0.034

1:5000

1.95^d1

1.88^bc1

1.83^d1

1.85^de1

0.52⁴

0.56⁴

0.74^b3

1.45^abc2

0.038

1:10,000

2.17^c1

2.01^b2,3

2.14^c1,2

1.96^cd3

0.60⁶

0.59⁶

1.68^a4

1.51^a5

0.047

40 µg

580

1:2500

2.00^d1

2.00^b1

1.82^d2

1.90^cde2

0.60³

0.58³

0.59³

0.70⁴

0.020

1:5000

2.10^cd1

2.11^b1

2.02^c1,2

1.95^cd2

0.58³

0.60³

0.55³

0.73⁴

0.033

1:10,000

2.34^bc1

2.12^b2

2.10^c2

2.02^cd2

0.65³

0.63³

0.60³

0.60³

0.034

600

1:2500

1.42^f1

1.37^f1,2

1.29^g2

1.42^g1

0.59³

0.62³

0.69³

0.63³

0.042

1:5000

1.58^e1

1.50^de1,2

1.42^f2

1.45^fg2

0.58³

0.61³

0.73⁴

0.72⁴

0.039

1:10,000

1.83^d1

1.65^d2

1.75^d1,2

1.53^f3

0.63⁴

0.59⁴

0.63⁴

0.75⁵

0.041

630

1:2500

1.42^f1

1.40^ef1

1.23^g2

1.41^g1

0.57³

0.58³

0.64³

0.65³

0.037

1:5000

1.56^e1

1.47^e1,2

1.42^f2

1.46^fg1,2

0.53³

0.57³

0.64³

0.75⁴

0.035

1:10,000

1.79^d1

1.63^d2,3

1.72^de1,2

1.59^f3

0.64⁴

0.64⁴

0.71⁴

0.72⁴

0.039

SEM

0.053

0.052

0.028

0.036

0.037

0.023

0.032

0.025