PerCP Conjugation Kit - Lightning-Link® (ab102907)

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Overview

  • Product name

    PerCP Conjugation Kit - Lightning-Link®

  • Product overview

    PerCP Conjugation Kit ab102907 uses a simple and quick process to conjugate an antibody to PerCP. It can also be used to conjugate other proteins or peptides.


    To conjugate an antibody to PerCP using this kit:
    - add modifier to antibody and incubate for 3 hrs
    - add quencher and incubate for 30 mins
    The conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.


    Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to PerCP.


     

  • Notes

    Amount and volume of antibody for conjugation to PerCP

     Kit size   Recommended maximum
    amount of antibody         		 Maximum antibody 
    volume1
    30 µg   3 x 10 µg 3 x 10 µL
    100 µg   100 µg  100 µL
    300 µg   3 x 100 µg  3 x 100 µL
    1 mg   1 mg 1 mL

    1 Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 5mg/ml or < 0.5 mg/ml should be diluted /concentrated.

     

    Buffer Requirements for Conjugation

    Buffer should be pH 6.5-8.5.

    Compatible buffer constituents 
    If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.

    50mM / 0.6% Tris1 0.1% BSA 50% glycerol
    0.1% sodium azide PBS Potassium phosphate
    Sodium chloride HEPES Sucrose
    Sodium citrate EDTA Trehalose

    1 Tris buffered saline is almost always ≤ 50 mM / 0.6%

    Incompatible buffer constituents

    Thiomerosal Proclin Glycine
    Arginine Glutathione DTT

    If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.

    Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture 

  • Tested applications

    Suitable for: Conjugationmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab102907 in the following tested applications.

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

Application Abreviews Notes
Conjugation Use at an assay dependent dilution.

Images

  • Conjugation - PerCP Conjugation Kit (ab102907)
    Conjugation - PerCP Conjugation Kit (ab102907)

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

Protocols

References

This product has been referenced in:

  • Bok S  et al. In vivo imaging of activated microglia in a mouse model of focal cerebral ischemia by two-photon microscopy. Biomed Opt Express 6:3303-12 (2015). Read more (PubMed: 26417502) »
See 1 Publication for this product

Publishing research using ab102907? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Question

Has 100 ug of a protein in 250 uL. How should protocol be modified? The first step says to add 10 uL of modifier to 100 uL of antibody- should 25 uL be added since 250 uL of antibody is used?

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Answer

Thank you for your call yesterday and for your patience while I have been in touch with the scientists in the lab regarding your enquiry.

My colleagues have pointed out that the protein should be concentrated for the best results. The entire 250 uL should not be added to the single 100 ug vial of conjugate even though the protein is less concentrated. The conjugation will not work with 250 uL of sample volume. If the protein can not be concentrated, you can try with the dilute protein but the 250 uL will need to be split over the multiple vials of conjugate, so that no more than 100 uL will be added per vial. This should give some conjugated product but please note that the results will not be optimal.

I hope this information will be useful, but please let me know if you have any further questions or need anything else regarding these kits, and I will be happy to help. Have a nice day.

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Question

Hello, I have another question concerning the PerCP/Cya5.5 reagent I got in the labeling kit. Could you tell me the amount (either estimation or measured) of Cya5.5 per PerCP molecule ? Thank you and best regards,

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Answer

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 this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

I ordered kits mentionned below. I would like to know if you have any data concerning the extinction coefficient of PerCP and PerCP-Cy55 at its optimal absorption wavelength. Thank you and best regards,

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Answer

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 coefficient (cm-1M-1)= unknown I hope this information is helpful. Please let me know if you have any questions.

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