The dilution factors suggested in the following table are presented in the form of a range because the optimal dilution is a function of many factors, such as antigen density, permeability, etc. The actual dilution used must be determined empirically, but should fall within the suggested ranges based on customer feedback and our firsthand experience. Dilutions outside these ranges may be required occasionally, and should be determined based on actual circumstances.
|
| Product |
ELISA |
Western
Blots |
Histo-/
Cyto-Chemistry |
Flow
Cytometry |
| Unconjugated secondary antibodies [IgG, F(ab')2, Fab] |
10-20 µg / ml |
| All DyLight and Cy3 Conjugates [IgG, F(ab')2, Fab] |
|
|
1:100-
1:800 |
1:100-
1:800 |
AMCA, Cy2, FITC, TRITC, RRX, and Texas Red Conjugates
[IgG, F(ab')2, Fab] |
|
|
1:50-
1:200 |
1:50-
1:200 |
| Cy3 Conjugates [IgG, F(ab')2, Fab] |
|
|
1:100-
1:800 |
1:100-
1:800 |
| Cy5 Conjugates [IgG, F(ab')2, Fab] |
|
|
1:100-
1:400 |
1:100-
1:400 |
| Phycoerythrin Conjugates [F(ab')2, Streptavidin] |
|
|
|
1:50-
1:200 |
| Horseradish Peroxidase Conjugates [IgG, F(ab')2] |
1:5,000-
1:100,000 |
1:5,000-
1:100,000
(non-ECL)
1:10,000-
1:200,000
(ECL) |
1:500-
1:5,000 |
|
| Alkaline Phosphatase Conjugates [IgG, F(ab')2] |
1:5,000-
1:50,000 |
1:5,000-
1:50,000 |
|
|
Biotin-SP Conjugates[IgG, F(ab')2]
(using Fluorophore-Conjugated Streptavidin) |
|
|
1:200-
1:1,000 |
1:200
1:1,000 |
Biotin-Sp Conjugates [IgG, F(ab')2]
(using enzyme-Conjugated Streptavidin) |
1:20,000-
1:400,000 |
1:20,000-
1:400,000 |
1:500-
1:5,000 |
|
| Horseradish Peroxidase-Conjugated Streptavidin |
1-2 µg / ml |
0.01-0.1
µg / ml |
1-2 µg / ml |
|
| Alkaline Phosphatase-Conjugated Streptavidin |
1-2 µg / ml |
0.1-1µg / ml |
1-2 µg / ml |
|
| All DyLight Conjugated Streptavidin |
|
|
0.5-2 µg / ml |
0.5-2 µg / ml |
| Cy3 Conjugated Streptavidin |
|
|
0.5-2 µg / ml |
0.5-2 µg / ml |
| Cy5 Conjugated Streptavidin |
|
|
1.0-4 µg / ml |
1.0-4 µg / ml |
| All Other Fluorophore Conjugated Streptavidin |
|
|
2-5 µg / ml |
2-5 µg / ml |
| 4 nm Gold-Antibody Complexes |
|
|
1:20-1:200 |
|
| 18 nm Gold-Antibody Complexes |
|
|
1:10-1:20 |
|
| 6, 12 nm Gold Antibody Complexes |
|
|
1:20-1:40 |
|
| Peroxidase-Anti-Peroxidase (PAP) |
|
|
25-50 µg / ml |
|
|
