"We only use secondary antibodies from Jackson ImmunoResearch whenever possible! Customer service is also excellent and the price point for their products is very competitive."
Elizabeth Soberg, University of Washington
|
Technical Service e-mail
tech@jacksonimmuno.com |
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. 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 whole IgG and F(ab') 2 secondary antibodies |
10-20 µg / ml
| |||
| Unconjugated Fab secondary antibodies |
20-40 µg / ml
| |||
| Most Alexa Fluor®, DyLight, and Cy3 Conjugates [IgG, F(ab') 2, Fab] |
1:100-
1:800 |
1:100-
1:800 | ||
| Alexa Fluor® 680 and 790 Conjugates | 1:50,000-200,000 | |||
|
AMCA, Cy2, FITC, TRITC, RRX, and Texas Red Conjugates
[IgG, F(ab') 2, Fab] |
1:50-
1:200 |
1:50-
1:200 | ||
| Cy5 Conjugates [IgG, F(ab') 2, Fab] |
1:100-
1:400 |
1:100-
1:400 | ||
| Phycoerythrin and Allophycocyanin Conjugates [IgG, F(ab') 2, Streptavidin] |
1:50-
1:200 | |||
|
PerCP Conjugates
[IgG, F(ab') 2, Streptavidin] |
1:25-
1:100 | |||
|
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 Alexa Fluor®-, DyLight 405-, and Cy3-Conjugated Streptavidin | 0.5-2 µg / ml | 0.5-2 µg / ml | ||
| Cy5-Conjugated Streptavidin | 0.5-2 µg / ml | 0.5-2 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 | |||
