概述

  • 产品名称
    PE / R-Phycoerythrin偶联试剂盒
  • 产品概述

    R-PE Conjugation Kit ab102890 uses a simple and quick process to conjugate an antibody to R-PE. It can also be used to conjugate other proteins or peptides.


    To conjugate an antibody to R-PE 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 R-PE.

  • 说明

    Amount and volume of antibody for conjugation to R-PE

     Kit size   Recommended 
    amount of antibody
    1 
    Maximum antibody 
    volume2
    30 µg   3 x 10 µg 3 x 10 µL
    60 µg   60 µg  60 µL
    180 µg   3 x 60 µg  3 x 60 µL
    600 µg   600 µg 600 µL

    The selling size of this product has been changed – it is now based on the amount of antibody that can be conjugated with the kit, not the amount of PE mix provided. The quantity of reagents provided have not changed or the amount of antibody that can be conjugated.

    1 Kit is designed to give a 1:1 molar ratio of antibody to PE after conjugation.

    2 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%/1% BSA2 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%
    2 1% BSA gives lower quality conjugates, BSA can also interfere with the use of the conjugated antibody in tissue staining.

    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 media are incompatible.

性能

图片

  • Flow cytometry histogram showing integrin beta-3 positive population of platelets from wildtype and knockout mice. Integrin beta-3 antibody was conjugated using EasyLink R-Phycoerythrin conjugation kit (ab102918). Flow cytometry was performed using platelets from wild type and integrin beta-3 knockout mice. Mice that expressed beta-3 (shown in red) had a clear shift in FL-2 fluorescence over beta-3 knockout mice (shown in black).

实验方案

文献

This product has been referenced in:
  • Boopathy GTK  et al. Cavin-2 regulates the activity and stability of endothelial nitric-oxide synthase (eNOS) in angiogenesis. J Biol Chem 292:17760-17776 (2017). Read more (PubMed: 28912276) »
  • McGinn OJ  et al. Targeting the 5T4 oncofetal glycoprotein with an antibody drug conjugate (A1mcMMAF) improves survival in patient-derived xenograft models of acute lymphoblastic leukemia. Haematologica 102:1075-1084 (2017). Read more (PubMed: 28341731) »
See all 5 Publications for this product

客户评价及客户问答

1-8 of 8 Abreviews or Q&A

Minimal handling, and it worked!!

Excellent Excellent 5/5 (Ease of Use)
Abreviews
The kit was extremely easy to use and effective despite the very small amount of antibody labeled (60 ug)- minimal handling, and it worked!!
Flow cytometry histogram showing integrin beta-3 positive population of platelets from wildtype and knockout mice. Integrin beta-3 antibody was conjugated using EasyLink R-Phycoerythrin conjugation kit (ab102919). Flow cytometry was performed using platelets from wild type and integrin beta-3 knockout mice. Mice that expressed beta-3 (shown in red) had a clear shift in FL-2 fluorescence over beta-3 knockout mice (shown in black).

Abcam user community

Verified customer

提交于 Oct 17 2013

Answer

Compounds containing amines (such as Tris) compete with the antibody or other protein for the R-Phycoerythrin. Unusually for an amine, Tris has little effect on conjugation efficiency as long as the concentration is 20mM or less. So, if 100 mM (0.1 M) is going to be the final concentration of the Tris, the conjugation may not work.

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Answer

Thank you for contacting us.

We have the same integrin beta 1 antibody [12G10] that is FITC conjugated as ab150002, just so you know, but we do not have an R-PE conjugated version.

But if you would like to conjugate the ab30394 to R-PE using our EasyLink kit ab102918, that would definitely work. Once the antibody is conjugated using the kit, it is ready to use and no further purification is necessary.

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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Answer

Thank you for contacting us.


I can confirm that the R-Phycoerythrin (RPE) conjugation kit is indeed free of animal components.


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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Answer

Thank you for contacting us. We have not tested ab75803 in Flow Cytometry and do not have any data regarding suitability in this application. You would have to purify the antibody before conjugating it to remove the BSA or the BSA may be conjugated.
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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Answer

Thank you for contacting us.

The EasyLink conjugations require antibody concentrations of at least 1mg/ml. Below that, the conjugation becomes inefficient and you risk wasting the conjugate, not to mention the antibody. For antibodies of lower concentrations, we recommend bringing them up to 1mg/ml or more with a concentration columns. We have two sizes: one column or three columns.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=102776).

Click here (or use the following: https://www.abcam.com/index.html?datasheet=102778).

These are very easy to use, and ensure effective conjugations. Please let me know if you have any questions.

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Answer

Thank you for contacting us with your question.

Ab102918 has been developed to conjugate antibodies, however it targets available amine groups, so it can conjugate proteins and peptides provided there is at least one aminegroup (lysine) available.

For this reaction to be successful, the same restrictions do apply. The protein should be purified and in an amine free buffer (to prevent conjugation of buffer constituents), and the concentration should be at least 0.5 mg/mL. Depending on the protein size, the conjugation ratio can be very different than what is stated on the datasheet. For instance, many small peptides can conjugate to one molecule of PE, whereas we would expect an antibody to PE ratio of 1:1.

I hope this information will be useful, but please let me know if you have further questions or if there is anything else that we can do for you and I'll be happy to help.

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Answer

Thank you for contacting us.

We have a PE- conjugated antibody against cytokeratin, catalogue item ab52460:

Click here (or use the following: https://www.abcam.com/index.html?datasheet=52460).

For collagen I, we have only one unconjugated antibody that has been tested in flow cytometry. We do, however, have a kit that enables one to easily conjugate the antibody to PE.

Collagen I - ab23446
EasyLink PE - ab102919 or ab102918

For gp100, we have unconjugated antibodies but neither is in a formulation that is compatible with the conjugation kits for FITC. What I recommend is using the unconjugated anti-gp100, ab117853, followed by FITC-conjugated anti-mouse IgG2b, ab98702, pre-adsorbed to remove reactivity with mouse IgG1 (which is the isotype of ab52460, anti-cytokeratin). This should allow you to apply both antibodies to the same sample without cross-reaction of the secondary anti-IgG2b with mouse IgG1.

Click here (or use the following: https://www.abcam.com/index.html?datasheet=117853).

Click here (or use the following: https://www.abcam.com/index.html?datasheet=98702).

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

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