Site Specific, Clean Simple Antibody Labelling
The oYo-Link is fast becoming a popular antibody labelling system. With a ‘Mix and Illuminate’ protocol that delivers covalent antibody conjugation in 2 hours, with only 30 seconds hands on time and because the binding is site specific, it is truly an antibody labelling system, rather than a general protein bioconjugation.
This means that oYo-Link:
- Will not label stabilising proteins in your antibody buffer.
- Will not add the conjugate in the antibody binding site, rendering the antibody ineffective.
- Can be used to label antibodies at lower concentrations – as little as 1ug diluted down to 50ug/ml
- Has a limited heterogenicity of reaction product; with each antibody having 0,1 or 2 labels.
- May not need purification to remove any excess labelling reagents, offering 100% antibody recovery. oYo-Link will not bind to proteins in down stream applications, and antibodies only if there is UV illumination.
- Compatible with all common buffers, including BSA, Tris & Azide.
- Check out the full buffer and antibody compatibility table below!
Method Ready Antibodies
Alpha Thera offer oYo-Link with a range of labels
Oligonucleotides
Send us your oligonucleotide sequence, for a custom oYo-Link oligo for your experiment, ready to conjugate to your antibodies of choice.
- Any sequence
- Any length
- Any modification
Enzyme - HRP
Epitope Tags
10 Epitope tags to choose from enabling simple multiplexing, even with antibodies from the same species. Choose from His12, DYKDDDDK, VSV-G, V5, S, S1, NWS, AU1, AU5 and HSV1.
Fluorescence
oYo-Surelight enables fast labelling with PE, APC, PE-Cy7, or Far Red-APC.
Click Chemistry
oYo Link is offered with Azide, DBCO, Tetrazine or Thiol functional groups, for when antibody labelling is just the start! Over 90% more efficient click reactions.
Drugs
To help develop ADC’s oYo-Link can label your antibody with VcMMAE, VcMMAF, DM1 or DM4. AS attachment is limited to 1 or 2 drug molecules, precise Drug -Antibody Ratios (DAR) can be maintained.
Biotin
Alternatively, 2BScientific can help you access Alpha Thera’s antibody labelling service
oYo-Link® Buffer Compatibility
oYo-Link® products are compatible with all common buffers, including those containing storage proteins (e.g. BSA) and Tris. As a result, you will likely not need to dialyze or filter your antibody prior to labeling. oYo-Link also works across a wide range of pH values. The high affinity interaction between oYo-Link and IgG even enables antibodies to be labeled directly in serum or hybridoma supernatant, and antibody concentrations can be as low as 50 µg/mL.
| Buffer Category | Specific Chemical | Compatibility |
|---|---|---|
pH | 4-10 | |
Buffer | Tris buffer | ✔ |
PBS buffer | ✔ | |
HEPES Buffer (<20 mM) | ✔ | |
Borate Buffer | ✔ | |
MOPS Buffer (<20 mM) | ✔ | |
MES Buffer (50-500 mM) | Reduces photo-crosslinking efficiency by 50% | |
Salts | Sodium citrate | ✔ |
Sodium Azide (<1%) | ✔ | |
Chelating reagent | EDTA (<2.5mM) | ✔ |
Buffer additives | Trehalose | ✔ |
Sugars | ✔ | |
L-Arginine | ✔ | |
Proclin | ✔ | |
Thimerosal | ✔ | |
Merthiolate | ✔ | |
Glycine | ✔ | |
Glycerol (=<50%) | ✔ | |
Gelatin (0.5%) | ✔ | |
Storage protein | BSA | ✔ |
Serum | Human | ✔ |
Bovine | ✔ | |
Ascitic Fluid | ✔ | |
Cell culture medium | DMEM, RPMI-1640 | ✔ |
Hybridoma supernatant | ✔ |
oYo-Link® Antibody Compatibility
oYo-Link® exhibits differences in its ability to covalently photo-crosslink various Immunoglobulins (Ig). This variability exists both between antibodies from different species and between different antibody subclasses from the same species. The table to the left gives an overview of the cross-linking efficiency of oYo-Link to different Ig host species and subclasses. Or view our video for more information.
| Host Species | lg Subclass | Crosslinking Efficiency |
|---|---|---|
Human | lgG1 | +++ |
lgG2 | +++ | |
lgG3 | +++ | |
lgG4 | +++ | |
lgA | - | |
lgM | - | |
Mouse | lgG1 (Separate Product) | ++* |
lgG2a | +++ | |
lgG2b | ++ | |
lgG2c | +++ | |
lgG3 | +++ | |
Rat | lgG1 | + |
lgG2a | - | |
lgG2b | - | |
lgG2b kappa | + | |
lgG2c | +++ | |
Goat | lgG | ++ |
Rabbit | lgG | ++/+++** |
Sheep | lgG | ++ |
Cow/Bovine | lgG | ++ |
Dog | lgG | ++ |
Pig/Porcine | lgG | +++ |
Guinea Pig | lgG | +++ |
Armenian Hamster | lgG | + |
Chicken | lgY | - |
Donkey | lgG | +++ |
Horse/Equine | lgG | +++ |
Monkey | lgG | +++ |
+++: Efficient Crosslinking ++: Moderate Crosslinking -: No Crosslinking
* Labeling mouse lgG1 requires the use of oYo-Link mlgG1 and is available as a separate product for all labels with the exception of oYo-Link Epitope Tag.
oYo-Link mlgG1 should only be used to label mouse lgG1. It is not compatible with other antibodies.
** Labeling Rabbit lgG has been shown to produce both moderate and efficient crosslinking efficiency.
Note: To label mouse IgG1 antibody, you must order the oYo-Link mIgG1 products, which will only label mIgG1 antibody.
Note: While crosslinking efficiency is usually consistent for antibodies from a specific species/host, some exceptions exist. Therefore, customers are encouraged to check the labeling efficiency of their antibody if labeling efficiency is critical for an experiment.
