Overview
- Product nameEasyLink AMCA Conjugation Kit (1 x 1mg AMCA)See all AMCA kits ...
- Product overview
EasyLink AMCA Conjugation Kit provides a simple and quick process to conjugate your primary antibodies with AMCA. AMCA is a blue fluorescent dye which has an excitation wavelength of 350nm and an emission wavelength of 445nm.
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.
- Notes
Pre-conugation considerations
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.
Conjugation reaction
- 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.
Disclaimer
Labeling of the antibody will not work if the conjugation blocks the active paratope. This situation is rare.
- Tested applicationsConjugation more details
Properties
- Storage instructionsStore at -20°C. Please refer to protocols.
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Components 1 mg EL-AMCA Mix 1 vial EL-Modifier reagent 1 vial EL-Quencher reagent 1 vial - Research Areas
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Alternative names
- Aminomethylcoumarin acetate
Applications
Our Abpromise guarantee covers the use of ab102845 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| Conjugation | Conjugation |
EasyLink AMCA Conjugation Kit (1 x 1mg AMCA) images
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Conjugation - EasyLink AMCA Conjugation Kit (1 x 1mg AMCA) (ab102845)This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
References for EasyLink AMCA Conjugation Kit (1 x 1mg AMCA) (ab102845)
ab102845 has not yet been referenced specifically in any publications.
