Overview

  • Product nameAMCA Conjugation Kit
  • Product overview

    Abcam's 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.

     

  • Notes

    Pre-conjugation 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 the kit 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 sizeAmount of antibodyVolume of antibody
    Up to 20 µg10-20 µg4-10 µl
    Up to 200 µg100-200 µg40-100 µl
    Up to 2 mg1-2 mg400-1000 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

    1. Before adding the antibody to the mix, add 1 µl of Modifier reagent to each 10 µl of antibody to be labelled. Mix gently.

     

    • Remove cap from vial of AMCA mix and pipette the antibody sample (with added 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 Quencher  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

  • Alternative names
    • Aminomethylcoumarin acetate

Applications

Our Abpromise guarantee covers the use of ab102846 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 Use at an assay dependent dilution.

AMCA Conjugation Kit images

  • This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.

Protocols

References for AMCA Conjugation Kit (ab102846)

ab102846 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"