EasyLink Cy3® Conjugation Kit (1 x 1mg Cy3®) (ab102872)
- Product nameEasyLink Cy3® Conjugation Kit (1 x 1mg Cy3®)See all Cy3 kits ...
- Product overview
EasyLink Cy3 Conjugation Kit provides a simple and quick process to conjugate your primary antibodies with Cy3. Cy3 is a yellow fluorescent dye which has an excitation wavelength of 548nm and an emission wavelength of 561nm.
The conjugated antibody can be used straight away in WB, ELISA, IHC etc
Learn more about buffer compatibility, protein/secondary antibody conjugation and labelling chemistry in our FAQs.
This kit is sufficient for 1 labeling reaction of 1-2mg of antibody.
The purified antibody to be labeled should ideally be in 10-50mM amine-free buffer (e.g. MES, MOPS, HEPES, PBS), pH range 6.5 to 8.5. If the buffer is more concentrated or outside this pH range contact the technical support team.
Common non-buffering salts (e.g. sodium chloride), chelating agents (e.g. EDTA), and sugars have no effect on conjugation efficiency. Azide (0.02 to 0.1%) and BSA (0.1 to 0.5%) have little or no effect. Glycerol up to 50% has no effect.
Avoid buffer components that are nucleophilic, as these may react with EasyLink chemicals. Primary amines (e.g. amino acids or ethanolamine) and thiols (e.g. mercaptoethanol or DTT) fall within this class (Note: Tris-based buffers should be avoided). If your buffer contains primary amines and/or thiols, you should consider using our Concentration and Purification Kits (ab102778 or ab102784).
Recommended amount and volume of antibody for optimal results
Pack size Amount of antibody Volume of antibody Up to 20 µg 10-20 µg 4-10 µl Up to 200 µg 100-200 µg 40-100 µl Up to 2 mg 1-2 mg 400-1000 µl
Antibody concentrations of 1-4 mg/ml generally give optimal results. For conditions outside the suggested ranges contact the technical support team.
- Before adding the antibody to the EasyLink mix, add 1 µl of EL-Modifier reagent to each 10 µl of antibody to be labelled. Mix gently.
- Remove cap from vial of EasyLink mix and pipette the antibody sample (with added EL-Modifier) directly onto the lyophilized material. Resuspend gently by withdrawing and re-dispensing the liquid once or twice using a pipette.
- Replace cap on the vial and leave standing in the dark at room temperature (20-25°C) for 3 hours. Conjugations can also be set up and left overnight; longer incubation times have no negative effect on the conjugate.
- After incubating for 3 hours (or more), add 1 µl of EL-Quencher FD reagent for every 10 µl of antibody used and mix gently. The conjugate can be used after 30 minutes. The conjugates do not require purification.
Storage of conjugates
Storage at 4°C is recommended for any conjugate. A preservative may be desirable for long-term storage. Other storage conditions may also be satisfactory. The best conditions for any particular conjugate must be determined by experimentation.
Labeling of the antibody will not work if the conjugation blocks the active paratope. This situation is rare.
This material is also subject to proprietary rights of GE Healthcare Bio-Sciences Corp. and Carnegie Mellon University and made and sold under License from GE Healthcare Bio-Sciences Corp. This product is licensed for sale only for research. It is NOT licensed for any other use. There is no implied license hereunder for any commercial use.
COMMERCIAL USE shall include:
1. Sale, lease, license or other transfer of the material or any material derived or produced from it.
2. Sale, lease, license or other grant of rights to use this material or any material derived or produced from it.
3. Use of this material to perform services for a fee for third parties.
If you require a commercial license to use this material and do not have one, please return this material, unopened to Abcam Plc of 330 Cambridge Science Park, Cambridge, CB4 0FL, and any money paid for the material will
- Tested applicationsConjugation more details
- Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 mg EL-Cy3 mix 1 vial EL-Modifier reagent 1 vial EL-Quencher reagent 1 vial
- Research Areas
Our Abpromise guarantee covers the use of ab102872 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
EasyLink Cy3® Conjugation Kit (1 x 1mg Cy3®) images
This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
References for EasyLink Cy3® Conjugation Kit (1 x 1mg Cy3®) (ab102872)
ab102872 has not yet been referenced specifically in any publications.