製品の概要

  • 製品名Biotin (type B) Conjugation Kit
    Biotin (type B) キット 製品一覧
  • 製品の概要

    Abcam's  Biotin (type B) Conjugation Kit provides a simple and quick process to conjugate your primary antibodies with Biotin. It has been optimized to produce conjugates for assays in which the biotinylated protein is captured by streptavidin immobilized on a surface (e.g., plates, nitrocellulose, magnetic beads etc).

    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.

     

  • 特記事項

    Amount and volume of antibody

    The volume of the antibody sample ideally should be in the range 40-100µl. Antibody concentrations of 1mg/ml or greater generally give the best results. The Biotin kit contains a stable form of biotin and is much more efficient than standard biotin kits, thus no purification of the conjugate is required.

    Protocol

    1. Before you add antibody to the Biotin (type B) mix, add 10µl of Modifier reagent for each 100µl of antibody to be labeled. Mix gently.
    • Remove the screw cap from the vial of Biotin (type B) mix and pipette the antibody sample (with added Modifier) directly onto the lyophilised material. Resuspend gently by withdrawing and re-dispensing the liquid once or twice using a pipette.

     

    • Place the cap back on the vial and leave the vial standing for 3 hours in the dark at room temperature (20-25°C). Alternatively, and sometimes more conveniently, conjugations can be set up and left overnight, as the longer incubation time has no negative effect on the conjugate.

     

    • After incubating for 3 hours (or more), add 1µl of Quencher reagent for every 10µl of antibody used. The conjugate can be used after 30 minutes.

     

    Storage of conjugates

    For any new conjugate, initial storage at 4°C is recommended. A preservative may be desirable for long-term storage. Other storage conditions (e.g. frozen at -70°C or stored at -20°C with 50% glycerol) may also be satisfactory. The best conditions for any particular conjugate must be determined by experimentation.

    Sample Buffer

    Ideally, the antibody to be labeled should be in 10-50mM amine-free buffer pH range 6.5 to 8.5. However, many buffers outside these limits of concentration and pH can be accommodated. Modest concentrations of Tris buffer are also tolerated.

    Amine-free buffers, including MES, MOPS, HEPES and phosphate are compatible if they are in the concentration range 10-50mM and have pH values in the range 6.5-8.5, as the addition of Modifier provides the conditions necessary for efficient conjugation. Common non-buffering salts (e.g. sodium chloride), chelating agents (e.g. EDTA), and sugars may be present, as they have no effect on conjugation efficiency. Azide (0.02-0.1%) has little or no effect. If the amine-free buffer is relatively concentrated and outside the pH range 6.6-8.5 you may need to add more Modifier for each 10µl of antibody. Excess Modifier is provided so that you can check the pH of the buffer after the addition of the modifier. Ideally the pH should be around 7.3-7.6, though efficient conjugation occurs anywhere between pH 6.8 and 7.8. Avoid buffer components that are nucleophilic, as these may react with the kit chemicals. If your buffer contains primary amines (e.g. amino acids, ethanolamine) and/or thiols (e.g. mercaptoethanol, DTT), you should consider using our Concentration and Purification Kits (ab102778 or ab102784). (Note: Unusually, for an amine, Tris has little effect on conjugation efficiency as long as the concentration is 20mM or less).

     

    Disclaimer

    Labeling of the antibody will not work if the conjugation blocks the active paratope. This situation is rare.

     

    The antibody to be labelled should be purified, in an appropriate buffer for conjugation and at a suitable concentration, as described in the protocol booklet. If not, consider using our antibody purification and concentration kits.

  • アプリケーション適用あり: Conjugationmore details

製品の特性

    アプリケーション

    Our Abpromise guarantee covers the use of ab102867 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    アプリケーション Abreviews 特記事項
    Conjugation Use at an assay dependent dilution.

    Biotin (type B) Conjugation Kit 画像

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

    プロトコール

    Biotin (type B) Conjugation Kit (ab102867) 使用論文

    ab102867 has not yet been referenced specifically in any publications.

    Product Wall

    Both kits are NH2-reactive but the chemistry is proprietary. There is no need to change the pH to get an efficient reaction; the buffers in the various components of the kit will ensure that the reaction pH is correct.

    The N-terminus (if acces...

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    Notre gamme de kits de conjugaison peut être utilisée pour coupler des protéines avec la biotine.

    Ces kits fonctionnent en ciblant les groupes aminés libres sur les biomolécules donc si votre protéine contient des groupes aminés lib...

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    Thank you for contacting us.

    I have good news and bad news for you:

    We unfortunately cannot offer your a sELISA system in which the detection is conjugated either with HRP or Biotin.

    The good news is that we indeed have m...

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    Thank you for your enquiry.

    The two kits (Biotin A and B) use the same spacer. In both kits, there are multiple biotin molecules attached to a larger scaffold molecule, which is then attached to the antibody. Unfortunately, the molecular wei...

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    Thank you for your inquiry.

    Unfortunately, this would be an untested application. We do not currently have information about this type of use with this product.

    We apologize for any inconvenience this may cause.

    Thank you for your kind message and your interest in our products.
    I can confirm that all our antibodies against cytokeratin 7 is unconjugated. I regret to inform you that currently we do not offer custom-made antibody service.
    However, we ha...

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    Vielen Dank für Ihre Anfrage. In der Tat haben wir einige polyklonalen HSP60- Antikörper in unserem Katalog verfügbar, unter anderem zum Beispiel ab105853: ein Kaninchen-polyklonaler, der gegen das humane Volllängenprotein gerichtet ist und für West...

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    Some FAQs about the conjugation kits: http://www.abcam.com/index.html?pageconfig=resource&rid=13156 and please see the datasheet of the kits for additional information. 1. Yes, these concentrations of azide and BSA should be fine. 2. The recommen...

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