製品の概要

  • 製品名PerCP/Cy5.5® Conjugation Kit
  • 製品の概要

    Abcam's PerCP/Cy5.5 Conjugation Kit provides a simple and quick process to conjugate your primary antibodies with PerCP/Cy5.5. PerCP/Cy5.5 is a tandem conjugate: PerCP (energy donor) has an excitation wavelength of 477nm and Cy5.5 (energy acceptor) has an emission wavelength of 694nm.

    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 amount of antibody used for labeling should be 100µg (on each reaction). The volume in which the antibody is added ideally should be 40-100µl. Antibody concentrations in the range 1-4mg/ml are therefore ideal. However, concentrations and volumes outside these suggested limits have also yielded excellent conjugates. For any new antibody, optimization of the ratio of antibody to PerCP/Cy5.5 is often worthwhile.

    Protocol

    • Before you add antibody to the PerCP/Cy5.5 mix, add 10µl of Modifier reagent for each 100µl of antibody to be labeled. Mix gently.
    • Remove the screw cap from the vial of PerCP/Cy5.5 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 often 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, storage at 4°C is recommended. 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.

    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.

      

    COMMERCIAL USE shall include:

    1. Sale, lease, license or other transfer of the mate

     

    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

  • 別名
    • Peridinin chlorophyll protein complex

アプリケーション

Our Abpromise guarantee covers the use of ab102911 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.

PerCP/Cy5.5® 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

PerCP/Cy5.5® Conjugation Kit (ab102911) 使用論文

ab102911 has not yet been referenced specifically in any publications.

Product Wall

The conjugation is effective within the range 1-4 mg/ml, and less effective ouside of it. So, you still may be able to obtain some signal with a conjugation of a 0.5mg/ml antibody but it will be relatively weak. We recommend concentrating the antibody,...

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ab72479:
Like most phycobiliprotein conjugates, the ratio of the fluor to the antibody (F/P) can vary slightly from lot to lot due to the nature of the conjugation chemistry and the methods used to characterize the final product, but it is typicall...

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Thank you for contacting us.

Unfortunately, I am unable to tell you the number of Cy5.5 molecules. The amount of Cy5.5 per PerCP in the PerCP/Cy5.5 conjugation kit has been optimised to give excellent performance.


I hope t...

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Thank you for contacting Abcam. Here are the details you requested. Ab102907 (PerCP ): Abs max=482nm, Emission Max= 677nm , Extinction coefficient (cm-1M-1)= 380,000 Ab102911(PerCP/Cy5.5): Abs max=482nm, Emission Max= 700nm , Extinction coeffi...

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Thank you for your response.
You could certainly use the credit for purchasing a different product which is in our catalogue.
Your credit note ID is 19429.
I am sorry that this antibody did not perform as stated on the datasheet. I have ...

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Thank you for getting back to me and for providing some further information. Your co-operation is much appreciated. I understand that there is no additive in the storage buffer which could interact with the conjugation reaction. I would like to reassur...

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Thank you for getting back to me and for confirming the correct product code number of the kit you are having difficulties. Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help...

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