Jackson Immuno Research Inc.
specializing in secondary antibodies and conjugates

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Polyclonal AffiniPure-VHH™ Secondaries

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New VHH Fragment Antibodies for High-Resolution Immunostaining

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Whole IgG Affinity-Purified Secondary Antibodies

"I have used a wide variety of secondaries and Jackson ImmunoResearch has consistently been the best. The fluorophores are bright and stable and their selective (x reactivity removed) secondaries have always shown species specificity in multiple labeling."

Janet Duerr, Ohio University

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Suggested Dilution Factors

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Suggested Dilution Factors

The dilution factors suggested in the following table are presented as ranges because the optimal dilution is a function of many factors, such as antigen density, permeability, etc. The optimal working dilution should be determined empirically for each application.

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Application
Product Conjugate ELISA Western Blots Histo- /Cyto chemistry Flow Cytometry
Whole IgG and F(ab')2 secondary antibodies Unconjugated 10 - 20 µg/ml
Fab secondary antibodies Unconjugated 20 - 40 µg/ml
Whole IgG, F(ab')2 and Fab fragment secondary antibodies Alexa Fluor® 488, 555, 568, 594, 647, DyLight™ 405, and Cy™3 1:100 - 1:800
Whole IgG secondary antibodies Alexa Fluor® 680 and 790 1:50,000 - 1:200,000
Whole IgG, F(ab')2 and Fab fragment secondary antibodies AMCA, BV421™, BV480™, Cy2, FITC, TRITC, RRX, and Texas Red 1:50 - 1:200
Whole IgG secondary antibodies Cy™5 1:100 - 1:400
Whole IgG, F(ab')2 secondary antibodies and Streptavidin R - PE and APC 1:50 - 1:200
Whole IgG, F(ab')2 secondary antibodies and Streptavidin PerCP 1:25 - 1:100
Whole IgG and F(ab')2 secondary antibodies Horseradish Peroxidase 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
Whole IgG and F(ab')2 secondary antibodies Alkaline Phosphatase 1:5,000 - 1:50,000 1:5,000 - 1:50,000
Whole IgG, F(ab')2, Fab and AffiniPure-VHH® Secondary Antibodies Biotin-SP (using Fluorophore-Conjugated Streptavidin) 1:200 - 1:1,000 1:200 - 1:1,000
Whole IgG, F(ab')2 and AffiniPure-VHH® Secondary Antibodies Biotin-SP (using enzyme-Conjugated Streptavidin) 1:20,000 - 1:400,000 1:20,000 - 1:400,000 1:500 - 1:5,000
AffiniPure-VHH® Secondary Antibodies Cy3, DyLight 405, Alexa Fluors® 1:200 - 1:800
AffiniPure-VHH® Secondary Antibodies Rhodamine Red-X 1:100 - 1:400
Streptavidin Horseradish Peroxidase 1 - 2 µg/ml 0.01 - 0.1 µg/ml 1 - 2 µg/ml
Streptavidin Alkaline Phosphatase 1 - 2 µg/ml 0.1 - 1 µg/ml 1 - 2 µg/ml
Streptavidin Alexa Fluors®, DyLight 405, and Cy3 0.5 - 2 µg/ml 0.5 - 2 µg/ml
Streptavidin Cy5 0.5 - 2 µg/ml 0.5 - 2 µg/ml
Streptavidin All Other Fluorophores 2 - 5 µg/ml 2 - 5 µg/ml
Whole IgG secondary antibodies 4 nm Gold-Antibody Complexes 1:20 - 1:200
Whole IgG secondary antibodies 6, 12 nm Gold-Antibody Complexes 1:20 - 1:40
Whole IgG secondary antibodies 18 nm Gold-Antibody Complexes 1:10 - 1:20
Whole IgG secondary antibodies, Streptavidin and ChromPure 40 nm Gold-Antibody Complexes 1:10 - 1:100
Peroxidase-Anti-Peroxidase (PAP) 25 - 50 µg/ml
FabuLight™ Anti-IgM To complex with primary antibody in solution, use 1:1 weight ratio of Fab: primary antibody (15:1 molar ratio).
FabuLight Anti-IgG To complex with primary antibody in solution, use 1:1 weight ratio of Fab: primary antibody (3:1 molar ratio).
Normal Serum 5% (v/v) for blocking
ChromPure® 10 µg/ml

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