Flow Cytometry with JIR

For flow cytometry we offer three fluorescent proteins (R-PE, APC, and PerCP) conjugated to many highly adsorbed secondary antibodies, streptavidin, and purified immunoglobulin controls. The table of Secondary Antibodies for Flow Cytometry lists all of the antibodies to which we conjugate these proteins. Also shown in this table are the same highly adsorbed antibodies conjugated to Biotin-SP and fluorescent dyes appropriate for flow cytometry (Alexa Fluor® 488, FITC, and Alexa Fluor® 647). Note that many antibodies listed elsewhere in tables of Whole IgG and F(ab’)2 Fragments also can be used for flow cytometry.

Excitation (left) and emission (right) spectra of Alexa Fluor® 488/FITC (green), R-PE (red), PerCP (blue), Alexa Fluor® 647 (black), and APC (brown). Peak heights were normalized after the spectra were obtained with an M-series spectrofluorometer system from Photon Technology International, Inc.

Fluorophore Excitation Peak Emission Peak (nm)
Alexa Fluor® 488 493 519
Fluorescein, FITC 492 520
R-Phycoerythrin R-PE many, 488 580
Allophycocyanin APC many, 650 670
Peridinin-Chlorophyll-protein, PerCP many, 488 675

Recommended dilution ranges for Flow Cytometry are shown in the table below.

Product Flow Cytometry
Unconjugated whole IgG and F(ab’) 2 secondary antibodies 10 – 20 µg / ml
Unconjugated Fab secondary antibodies 20 – 40 µg / ml
All Alexa Fluor®, DyLight, and Cy3 Conjugates [IgG, F(ab’) 2, Fab] 1:100 – 1:800
AMCA, Cy2, FITC, TRITC, RRX, and Texas Red Conjugates [IgG, F(ab’) 2, Fab] 1:50 – 1:200
Cy5 Conjugates [IgG, F(ab’) 2, Fab] 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
Biotin-SP Conjugates [IgG, F(ab’) 2] (using Fluorophore-Conjugated Streptavidin) 1:200 – 1:1,000
All Alexa Fluor®-, DyLight 405-, and Cy3-Conjugated Streptavidin 0.5 – 2 µg / ml
Cy5-Conjugated Streptavidin 0.5 – 2 µg / ml
All Other Fluorophore-Conjugated Streptavidin 2 – 5 µg / ml

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