oYo-Link® DNP

FOR RESEARCH USE ONLY: Site-specific labeling of compatible antibodies with up to two labels of DNP for ELISA, flow cytometry and immunofluorescence.

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oYo-Link®DNP Antibody Labeling Reagents

For research use only.

oYo-Link® DNP enables site-specific labeling of compatible antibodies with up to two labels of DNP, supporting high specificity and amplified detection in assays such as ELISA, Western blotting, and immunohistochemistry.

Simply mix oYo-Link® DNP with your antibody, followed by photocrosslinking using non-damaging black light (365 nm) for a total of 2 hours.  Photocrosslinking can be carried out in most buffers, including those containing amine-containing molecules (e.g., TRIS, glycine) and storage proteins (see the Buffer Compatibility Table here).

 

DNP-antibody conjugates offer robust, well-characterized hapten-based detection with high sensitivity, reproducibility, and broad assay compatibility, making them widely used in immunodetection and assay development. 

Because DNP exhibits no endogenous presence in biological samples, the resulting conjugates deliver exceptionally low background and high signal-to-noise performance.

Detection is achieved using anti-DNP antibodies, which are widely available and can be paired with multiple reporter systems (e.g., HRP, alkaline phosphatase, or fluorescent labels), enabling flexible readout formats including colorimetric, chemiluminescent, and fluorescent assays. 

These features make DNP conjugates particularly useful in ELISA, flow cytometry, immunofluorescence, and assay standardization workflows.

 

Requirements: To label your compatible antibodies with oYo-Link®, you will need a light source emitting at 365nm (6-10W). Purchase AlphaThera’s LED Photo-Crosslinking Device here, or use a compatible device.

Placing an order: For credit card purchases, proceed with checkout on website. For bulk order or to request a quote click here. 

Technical Support: For any technical questions please contact our technical support team here. 

oYo-Link DNP product pack image

oYo-Link® Protocol - Quick Reference Guide

Download this oYo-Link® Protocol/Quick Reference Guide for instructions on how to site-specifically label your antibody with oYo-Link®.

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Video: FAQ - What is the labeling efficiency of oYo-Link®?

FAQ: What is the conjugation efficiency of oYo-Link® Site Specific Antibody Labeling Reagents? For most subclasses and species of antibody, oYo-Link® will result in conjugation of 2 labels per antibody, therefore achieving approximately 100% labeled IgG. However, there are a few antibodies where the conjugation is slightly less efficient.

 

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Antibody Conjugation User Manual

Read our antibody conjugation usual manual for instructions on how to easily conjugate your antibody with oYo-Link products.

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Demo: oYo-Link® Antibody Conjugation

Find out how to site-specifically label your antibody with just two steps: mix and illuminate. 30 seconds hands-on time, 2 hours total.

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oYo-Link® Product Brochure

Download our brochure for more information and to access 20% off your first order of oYo-Link® Antibody Labeling products.

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Figure 1. BSA detection via ELISA using anti-BSA antibodies conjugated with oYo-Link DNP.

Plates were coated with BSA at varying concentrations (log scale, ng/mL), with binding measured by absorbance at 450 nm. After the addition of the oYo-DNP-conjugated anti-BSA rabbit antibody (circles), the oYo-Link Single Biotin-anti-DNP Goat antibody (Bethyl Laboratories, A150-117A with 1:5000 dilution) was applied, followed by SA-HRP (Thermofisher, N100). The resulting data demonstrates a dose-dependent sigmoidal response that reaches saturation at elevated antigen levels. Specificity is confirmed by the oYo-DNP-conjugated rabbit isotype control (squares), which produced minimal signal.

Specifications:

Product Name

oYo-Link® Dinitrophenol (DNP)
Catalog #

AT4003
Lot #

See Product Spec Sheet shipped with product.
MSDS

Download the MSDS here.
Antibody and Buffer Compatibility

View the oYo-Link Antibody & Buffer Compatibility Table here.

Note: To label mouse IgG1 antibody, you must order the oYo-Link mIgG1 products, which will only label mIgG1 antibody.
Molecular Weight

8 kDa
Concentration

270 µg/mL after resuspension
Molar Concentration

33 µM after resuspension
Storage Buffer

1 x PBS, pH7.3 after resuspension
Shipping Condition

This product is shipped as a clear or white pellet at ambient temperature.
Reconstitution

Before opening, briefly centrifuge each tube to ensure that the entire pellet is at the bottom. Add 200 μL sterile ddH2O and vortex tube briefly until fully dissolved.
Storage and Stability

The dried product can be stored at 4°C or -20°C upon receipt. Following reconstitution, the product can be stored at 4°C for short-term durations (< 1 week) and -20°C to -80°C for longer time-spans.
Conjugation Instructions

After reconstitution, each tube is capable of labeling 200 µg of compatible antibody. Please find and follow the Antibody Conjugation User Manual and Antibody Immobilization Protocol at https://alphathera.com/user-manuals for instructions.
Other Notes

Disposal of product should be performed in compliance with all applicable federal, state, and local regulations. This product does not meet hazard classification criteria based on evaluations made by our company under the OSHA Hazard Communication Standard, 29 CFR 1910.1200.

Will oYo-Link work with my antibody?

Please check the antibody compatibility table. Note: oYo-Link mIgG1 can be used to label mouse IgG1 antibody. Currently, oYo-Link mIgG1 is only available for certain products. Please check the product webpage. oYo-Link mIgG1 only labels mouse IgG1 and is not compatible with other antibodies.

Can oYo-Link be used to label antibodies in the presence of albumin and/or Tris?

Yes, since oYo-Link specifically bind to the heavy chain of IgG, it works in the presence of both albumin and Tris. oYo-Link is compatible with all common buffers. See the full buffer compatibility table here.

If oYo-Link DNP has accidentally been left at room temperature for a week, will it still work?

With the exception of oYo-Link® DBCO, oYo-Link® Thiol, and oYo-Link® SureLight® APC, Far-Red APC, PE, and PE-Cy7, all oYo-Link products are shipped as white or clear pellets and are very stable. While we recommend cold storage, they will not exhibit any loss of activity if left at room temperature for a week.

oYo-Link DBCO must be shipped and stored cold (-20°C) immediately upon arrival, otherwise there will be a loss in activity.

oYo-Link® SureLight® APC, Far-Red APC, PE, and PE-Cy7 are shipped as liquid with cold gel packs and should be stored at 2-8°C in the dark. Do not freeze.

Does AlphaThera offer a conjugation service?

Yes. We offer a conjugation service for $75 per antibody plus the cost of reagents. Purification (FPLC) is available for $250 per conjugate (please note that bulk orders may incur an additional charge).
You can find out more about our conjugation service here. For conjugation services, you will need to send us your antibody.

How many oYo-Link labels will be conjugated to each antibody?

For most subclasses and species of antibody, oYo-Link will result in the conjugation of 2 labels per antibody (maximum), one label per heavy chain. For example, ~95% of human IgG and Rabbit IgG will have two labels per antibody. However, there are a few antibodies such as mouse IgG2b and goat IgG, where the conjugation is slightly less efficient. In this case, 60 to 80% of the antibody will be labeled and have a mixture of 0, 1 and 2 labels per antibody. See the antibody conjugation efficiency table here.

How will I know if oYo-Link has been successfully conjugated to my antibody?

Antibody conjugation can be checked on SDS-PAGE gel. An example is present in the supporting data on each product page.

Does 365nm UV illumination damage the antibody and its binding affinity?

The conjugated and unconjugated antibody binding affinities have been tested via both ELISA and cell binding assays and don’t show any difference.

Do I need to remove free oYo-Link after conjugating to antibody?

For most immunoassay applications, purification is not necessary. In our standard recommended protocol, the molar ratio of oYo-Link-to-antibody is 5:1. These conditions ensure that the antibody conjugation efficiency reaches its maximum; however there will be a slight excess remaining in your antibody mixture. This is usually removed during immunoassay washing steps. If your application requires high antibody-conjugate purity, our oYo-Capture Magnetic Beads can be used to quickly remove free oYo-Link after antibody conjugation. Please note that oYo-Capture™ magnetic beads cannot be used for any secondary anti-Rabbit IgG antibody conjugated with an oYo-Link product. If you need to purify your oYo-Link conjugated Rabbit IgG antibody, please don’t hesitate to contact us for a detailed protocol.

What is the lowest amount of antibody that can be labeled by oYo-Link reagents?

We recommend working with at least 1 µg of antibody; however, it is possible that even lower amounts of antibody can be labeled.

Can antibodies that are highly dilute be labeled by oYo-Link reagents?

Yes, antibodies can be efficiently labeled even if diluted to concentrations as low as 50 µg/mL.

Can we order directly from you instead of your distributors?

Both options are feasible. You may place the order either through us or through our distributors. View our list of distributors here.

Can you conjugate a peptide to an antibody?

Yes, we also offer protein-antibody conjugation; however, this would be handled as a custom order. Our approach involves a click-chemistry reaction to attach the protein of interest to oYo-Link®, followed by site-specific crosslinking to the antibody. Please contact us to discuss your project.

oYo-Link® DNP Advantages

  • Enhanced assay sensitivity – Hapten-based signal amplification enables sensitive detection of low-abundance targets.
  • Low background, high signal-to-noise ratio – DNP exhibits no endogenous presence in biological samples, enabling highly specific detection.
  • Flexible detection formats – Compatible with a wide range of anti-DNP detection reagents for colorimetric, chemiluminescent, and fluorescent assays.
  • Site-Specific & Covalent Reaction – Covalent labeling of an antibody’s Fc region only, with no impact on antibody binding, ensures experimental reliability and reproducibility. 
  • <30 seconds “hands-on” time, 2 hours total. Two simple steps – mix and illuminate. 
  • Label as little as 1 µg of antibody per reaction.
  • Label diluted antibody solutions, as low as 50 µg/mL.
  • Compatible with all common buffers, including BSA, Tris & Azide. 
  • >90% labeling efficiency of most antibodies. 
  • After resuspension, aliquot and save for future reactions.

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