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Custom Antibody Conjugation Service

At Fortis Life Sciences, we offer a wide range of antibody conjugation solutions to enable you to find the perfect solution for your research purposes. We offer a fixed product range of conjugated antibodies, and also offer a custom antibody conjugation service to meet your specific experimental requirements.


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Antibody Conjugation Detection Methods

Antibodies conjugated with fluorophores, biomolecules, or enzymes are essential to both direct and indirect detection methods in immunoassays. Every immunoassay-based experiment requires specific type of labels/conjugates: for example, flow cytometry involves fluorescent labeled antibodies, while horseradish peroxidase (HRP)-labeled antibodies are useful for chemiluminescence based detection in Western blot analysis.
In immunoassays, the method of detection under consideration will determine whether a conjugated primary or secondary antibody is required. The direct and indirect methods are summarized below:

Direct Detection Method

One-step immunolabeling procedure in which a conjugated primary antibody binds its target antigen or analyte for generating a detection signal in the sample being tested.

Pros: Low background signal, minimal species cross-reactivity, great for multiplexing

Cons: Lower sensitivity, less flexibility from limited availability of conjugated primary antibodies


flow cytometry using dylight conjugated conjugated antibody

Indirect Detection Method

Two-step method in which an unconjugated primary antibody binds the target antigen or analyte, and then a labeled secondary antibody detects the bound primary for generating the final signal.

Pros: More flexible, highly sensitive (i.e., the signal is enhanced with secondary antibody)

Cons: Longer procedure, non-specific background of secondary limits multiplexing

flow cytometry using dylight conjugated conjugated antibody

Custom Conjugated Antibodies

The adoption of directly conjugated primary antibodies for use in multiplex immunoassays is increasing in research as well as diagnostic applications. Finding pre-made primary antibody conjugates for all of the protein targets and/or labels and trusting the quality/shelf-life of those conjugates can be challenging
To overcome this problem, we offer a custom conjugation service for our primary and secondary antibodies. Simply contact us with a catalog number of your unconjugated antibody with the specifications of a conjugated product that would fit your needs and our team will take care of the rest!
At Fortis, we offer custom conjugations with a variety of fluorophores including the DyLight®️ fluorophores (ranging from DyLight®️ 350 to DyLight®️ 800), as well as enzyme-conjugated antibodies and signal amplifying conjugates.

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DyLight® & Other Fluorophores

Fluorophores – also called fluorescent probes, fluorescent dyes, or fluorochromes – are proteins, organic small molecules, or synthetic polymers which are capable of absorbing light at a specific wavelength and then emitting the same at much longer wavelength(s). They have become essential to the mainstream immunoassay techniques in modern science, especially in microscopy and flow cytometry.
One of the key benefits of using fluorophore-based immunoassays is that they enable the detection of antigens expressed in very low abundance. Additionally, they exhibit differential/unique spectral properties which allows for simultaneous detection of multiple signals, i.e. fluorophores enable multiplexing options in immunoassays.
The DyLight® fluorophores are a series of fluorescent molecules exhibiting fluorescence emission covering nearly the entire visible light spectrum.
DyLight® exhibit higher or comparable fluorescence intensities and greater photo-stabilities versus their analogous Alexa Fluor®, CyDye™ and Li-COR® dyes. Wide Stokes shifts, narrow absorption/emission spectra, and an excellent aqueous solubility are a few other highlights of DyLight® fluorophores
Importantly, antibodies labeled with DyLight® have good aqueous solubility and stability over a broad pH (pH 4.0–9.0) which makes them ideal for multiplexing in cell-free as well as cell-based immunoassays.

DyLight® Conjugated Antibodies Applications

See examples of our DyLight® conjugated antibodies in action:

Flow Cytometry

flow cytometry using dylight conjugated conjugated antibody

Detection of CD3E by flow cytometry using anti-CD3E recombinant mAb (shaded red) [clone BL-298-5D12] [A700-016] and a DyLight® conjugated Goat-anti Rabbit IgG secondary antibody [A120-201D5].


IHC

IHC using dylight conjugated conjugated antibody IHC using dylight conjugated conjugated antibody

Detection of CD3E by flow cytometry using anti-CD3E recombinant mAb (shaded red) [clone BL-298-5D12] [A700-016] and a DyLight® conjugated Goat-anti Rabbit IgG secondary antibody [A120-201D5].


Western Blot

Multiplex fluorescent western blot

A fluorescent Western blot of hnRNP showing the multiplex use of our DyLight® 680 Conjugated Donkey anti-Mouse IgG-Heavy and Light Chain Cross-Adsorbed secondary antibody (red) [A90-337D6], with two other fluorescent labeled antibodies.



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Download fluorophore flyer
visible spectrum

The spectral characteristics of DyLight® and other fluorophore options we offer*:

Fluorescent Conjugate

Excitation Wavelength (nm)

Emission Wavelength (nm) 

Emission Color

Spectrally Equivalent Dyes

Cy2®

483-485

500

Green

Alexa Fluor® 488, ATTO® 488, CF®488A, DyLight® 488, FITC, Fluorescein

Cy3®

554

566

Yellow

Alexa Fluor® 555, CF®555, DyLight® 550, TRITC

Cy5®

649

666

Red

Alexa Fluor® 647, ATTO® 647N, CF®640R, CF®647, DyLight® 650

Cy5.5®

672-673

690

Red

Alexa Fluor® 680, CF®680, DyLight® 680, IRDye® 680

DyLight® 350

351

434

Blue

Alexa Fluor® 350, AMCA, CF®350

DyLight® 488

492

519

Green

Alexa Fluor® 488, ATTO® 488, CF®488A, Cy2®, FITC, Fluorescein

DyLight® 550

553

569

Yellow

Alexa Fluor® 555, CF®555, Cy3®, TRITC

DyLight® 594

591

617

Orange

Alexa Fluor® 594, ATTO® 594, CF®594, Texas Red

DyLight® 650

651

673

Red

Alexa Fluor® 647, ATTO® 647N, CF®640R, CF®647, Cy5®

DyLight® 680

676

705

Red

Alexa Fluor® 680, CF®680, Cy5.5®, IRDye® 680

DyLight® 755

750

771

Near Infrared

Alexa Fluor® 750, CF®750, Cy7®, APC, IRDye® 750

DyLight® 800

770

798

Infrared

CF®770, IRDye® 800

FITC

495

519

Green

Alexa Fluor® 488, ATTO® 488, CF®488A, DyLight® 488, FITC, Fluorescein

PE

565

576

Red

Alexa Fluor® 568, ATTO® 565, CF®568
*DyLight® is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries. We offer DyLight® antibody conjugates which we make at our antibody facility and validate in our in-house QC lab.

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Enzyme Conjugated Antibodies

Horseradish peroxidase (HRP) and alkaline phosphatase (ALP) are the two most common enzymes conjugated to antibodies for immunodetection applications. HRP and ALP act on chromogenic substrates to catalyze chemical reactions which produce insoluble, visible precipitates.
The quantity of chromogen produced from these reactions can serve as an indicator of the amount of antigen detected in the immunoassay. Additionally, the chromogen deposition in IHC/ICC can provide spatial information to determine localization of antigens within the tissue/cells. The benefits of chromogenic based detection include stability of signal, low cost, and integration into many visual analysis systems such as light microscopy and spectrophotometry.
Chromogenic detection with enzyme labeled antibodies is useful for ELISA, IHC/ISH, and Western blot immunoassays.

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IHC-P of human tonsil with rabbit anti-CD8 alpha recombinant mAb [clone BLR044F], A700-044 and HRP conjugated anti-rabbit secondary antibody followed by DAB-based detection

IHC-P of human tonsil with rabbit anti-CD8 alpha recombinant mAb [clone BLR044F] [A700-044] and HRP conjugated anti-rabbit secondary antibody followed by DAB-based detection.

Signal Amplifying Conjugates

In several types of immunoassays, the detection of low abundance target antigens can be facilitated using signal amplifier labeled antibodies, with biotin perhaps the most used biomolecule for signal amplification in immunoassays.
Multiple biotin molecules can bind to an antibody during the conjugation process, and when employed in an immunoassay, four molecules of fluorophore- or enzyme-labeled streptavidin bind to each of the biotin molecules at the detection stage. Accordingly, the biotin-streptavidin combination significantly enhances the detection signal.

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Request information for Custom Conjugation Service

Please fill the form below and use its "Notes" section to describe your custom conjugation requirements.
You can provide information on the catalog number of our unconjugated product to be conjugated, desired quantity (mg), the type of label (e.g., DyLight®, Biotin, HRP, ALP, etc.), buffer formulation, desired concentration, special custom QC, custom vialing and any documentation requirements (e.g., CofA).
Once this form is submitted, one of our experts will help you with a custom conjugation quote.