Technical Centre
TechFAQ #5

What are the suggested dilution factors to be used
for Jackson ImmunoResearch products?

Technical Service E-mail

tech@jireurope.com
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, 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  

NEWS!
   

          Jackson ImmunoResearch now offers selected Alexa Fluor® dye conjugates

 
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