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Fluorescent Dyes
Brilliant Violet™
421 & 480
- Combine with AF488, RR-X, & AF647 for 5-color imaging
- Compatible with commonly used filter sets
- Switch nuclear stain from DAPI to DRAQ5™ for additional labeling options
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Whole IgG Affinity-Purified Secondary Antibodies
"Without question, I can always turn to Jackson ImmunoResearch secondary antibodies for ECL detection of any primary antibody! They produce great signal at low dilutions."
W Thompson, OSHU
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Suggested Dilution Factors
|
Technical Service e-mail
tech@jacksonimmuno.com |
TechFAQ #5
What are the suggested dilution factors to be used for Jackson ImmunoResearch products?
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 | |||
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