Jackson Immuno Research Inc.
specializing in secondary antibodies and conjugates

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Multiple Labeling

With BV421™ & BV480™

  • Combine with AF488, RR-X, and 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

"We only use secondary antibodies from Jackson ImmunoResearch whenever possible! Customer service is also excellent and the price point for their products is very competitive."

Elizabeth Soberg, University of Washington

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Conjugate Selection

Technical Service e-mail

Probes Conjugated to Affinity-Purified Antibodies and Other Proteins

Technical Information on Enzymes and Biotin

Biotin-SP (long-spacer)

Enzymes - Horseradish Peroxidase and Alkaline Phosphatase

Technical Information on Fluorescent Dyes

Selection of fluorophores depends on:

  1. Instrument set-up. Examples include availability of light sources, filter sets, and detection systems.
  2. Degree of color separation desired for multiple labeling. For example, to achieve good color separation from Alexa Fluor® 488, choose a shorter wavelength-emitting fluorophore such as DyLight 405 and/or longer wavelength-emitting fluorphores such as Rhodamine Red-X, Alexa Fluor® 594, or Alexa Fluor® 647.
  3. Sensitivity required. For example, Alexa Fluor® 488 is brighter than FITC.

The following seventeen fluorescent dye conjugates (Figure 1, table 1) are currently available from Jackson ImmunoResearch, and includes dyes in the Alexa Fluor®, Brilliant Violet™ and Cyanine ranges. They cover the most commonly used excitation sources and filter sets from blue to infrared emissions. Click on each fluorophore for more information.

Please note that we also have limited stock of DyLight™ 488 / 549 / 594 / 649 and Texas Red® conjugates.

Effective 4-color imaging with good color separation, good photostability, and high sensitivity is possible with combinations such as DyLight 405, Alexa Fluor® 488, Rhodamine Red-X, and Alexa Fluor® 647.

Fluorophore Excitation Peak Emission Peak (nm)
DyLight 405 400 421
Brilliant Violet 421™ 407 421
Aminomethylcoumarin, AMCA 350 450
Brilliant Violet 480™ 436 478
Cyanine, Cy2 492 510
Alexa Fluor® 488 493 519
Fluorescein, FITC/DTAF 492 520
Indocarbocyanine, Cy3 550 570
R-Phycoerythrin R-PE many, 488 580
Rhodamine Red-X, RRX 570 590
Alexa Fluor® 594 591 614
Allophycocyanin APC many, 650 660
Alexa Fluor® 647 651 667
Indodicarbocyanine, Cy5 650 670
Peridinin-Chlorophyll-protein PerCP many, 488 675
Alexa Fluor® 680 684 702
Alexa Fluor® 790 792 803

Download text Files for all fluorescent dye conjugates to compare excitation and emission spectra

Table 1. Approximate peak wavelengths of excitation and emission for all fluorophore-conjugated, affinity-purified secondary antibodies offered by Jackson ImmunoResearch in order by increasing peak of emission. Approximate values are given for purposes of comparin one fluorophore with another. Actual values may vary depending on the spectrophotometer used in each laboratory.
Figure 1. Excitation (left) and emission (right) spectra of all fluorophores offered by Jackson ImmunoResearch. The conjugates in order of increasing wavelength of fluorescence are DyLight 405 (violet), AMCA (dark blue), Cy2 (light blue), Alexa Fluor® 488 (blue-green), FITC (hidden behind Alexa Fluor® 488), Cy3 (yellow), R-PE (dark pink, dotted), Rhodamine Red-X (orange), Alexa Fluor® 594 (red), Allophycocyanin (pink, dotted), Cy5 (green), Alexa Fluor® 647 (purple-hidden behind Cy5), PerCP (lavender), Alexa Fluor® 680 (gray), and Alexa Fluor® 790 (black).

Figure 1 illustrates the relative shape and position of each fluorophore in the peak region of its excitation and fluorescence emission following conjugation to antibodies. Quantitative comparisons should not be made since peak heights have been normalized. All spectra were obtained with an M-Series spectrophotometer system from Photon Technology International, Inc.

Technical Information on Colloidal Gold

LM Grade 4 nm and EM Grade 6 nm, 12 nm, and 18 nm Colloidal Gold - Antibody Complexes