TechFAQ #5

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

Technical Service E-mail

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. which we cannot predict. 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, however, and should be determined based on actual circumstances.

Product	ELISA	Western Blots	Histo-/ Cyto-Chemistry	Flow Cytometry
Cy2 and Conventional Fluorophore 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
Cy3 and 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