HTRF MAB ANTI MBP-EU - 5K PTS

HTRF MAB ANTI MBP-EU - 5K PTS

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品牌: Revvity
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    产品介绍
    产品信息

    Overview

    MAb Anti MBP-Eu cryptate is an IgG1 raised against a purified recombinant of Maltose Binding Protein (MBP) fusion protein labeled with Eu. It recognizes native as well as denatured-reduced forms of purified MBP or MBP fusion proteins.

    This reagent can be used in both biochemical and cellular formats to study a wide variety of interactions: protein/protein, protein/peptide, protein/DNA, protein/RNA, protein/carbohydrate, protein/small molecule, receptor/ligand.

    HTRF can detect a broad range of affinity constants ranging from picomolar to low millimolar.

    Specifications

    Assay Points
    20000
    Assay Target Type
    Fluorescent reagent
    Assay Technology
    HTRF
    Brand
    HTRF
    Quantity
    1
    Therapeutic Area
    Cardiovascular
    Infectious Diseases
    Inflammation
    Metabolism/Diabetes
    NASH/Fibrosis
    Neuroscience
    Oncology & Inflammation
    Rare Diseases
    Unit Size
    20,000 Assay Points

    Video gallery

    How it works

    Assay principle

    In an HTRF interaction assay, one partner is labeled (directly or indirectly) with the donor, and the other with the acceptor (again, directly or indirectly). The intensity of the signal is proportional to the binding of the 2 partners. In the example shown here: MAb Anti MBP-Eu cryptate binds to the MBP tagged partner A while partner B* binds to a specific Ab labeled with an HTRF acceptor. *partner B can also be biotinylated, tagged, Fc fused. In these cases, use the corresponding HTRF reagent (anti-Tag, anti-species, protA, Streptavidin) labeled with acceptor for the detection.

    HTRF MAb Anti MBP-Eu cryptate 1.svg

     

    Assay protocol

    The example on the right describes the protocol using a 20 µL final assay volume for detecting an interaction between a MBP-tagged partner A and a non-tagged partner B*. Dispense the 2 partners (10 µL), incubate, add MAb Anti MBP-Eu cryptate (5 µL) and anti-partner B labeled with acceptor (5 µL), incubate and read. *partner B can also be biotinylated, tagged, Fc fused or directly labeled. In these cases, use the corresponding HTRF reagent (anti-Tag, anti species, protA, Streptavidin) labeled with acceptor for the detection.

    HTRF MAb Anti MBP-Eu cryptate 2.svg

     

    Assay details

    How do the number of tests relate to active moiety?

    The average conjugate quantity per well reflects overall biological material content. Using the active moiety amount is generally preferred to the quantity of total conjugate. For Cryptate and d2 conjugates, the total conjugate amount equals that of the active moiety, since the molecular weight of the label is negligible. This is not the case for XL665 labeled entities for which the quantity of total conjugate will vary depending on the final molar ratio of the XL665 conjugate, however, the amount of active moiety, provided by Revvity, is constant and based on the number of tests ordered.

    HTRF MAb Anti MBP-d2 3.svg

     

    Recommended quantities of Cryptate and XL665 conjugates 

    Cryptate conjugates must not be excessive in order to prevent reader saturation and an unacceptable level of background. In most cases, a cryptate concentration of 1 to 5nM is appropriate, and will generate 20,000 to 80,000 cps at 620 nm depending on the HTRF compatible reader used. The XL665 conjugate must match its assay counterpart as closely as possible in order for the maximum number of biomolecules to be tagged with the XL665 acceptor. Thus, to detect a tagged molecule at an assay concentration of 20nM, the concentration of anti-Tag-XL665 should be equimolar or higher.

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    货号:
    61MBPKAA
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