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Fluorescent TrueBlot®: Anti-Rabbit IgG Fluorescein Accessory Reagents

Catalog No. ABIN6698841
$213.00
Plus shipping costs $45.00
100 μL
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  • Target Name (Antigen)
    Reactivity
    Rabbit
    Host
    Mouse
    Conjugate
    FITC
    Application
    FLISA, Flow Cytometry (FACS), Fluorescence Microscopy (FM), Immunohistochemistry (IHC), Western Blotting (WB)
    Brand
    TrueBlot®
    Characteristics

    Synonyms: FITC, TrueBlot, FITC TrueBlot ULTRA, Fluorescein TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, Fluorescent TrueBlot, Rb TrueBlot

    Background: Rabbit IgG TrueBlot® is a unique fluorescein conjugated Anti-rabbit IgG immunoblotting (second step) reagent. Rabbit IgG TrueBlot® enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Fluorescent Western Blot data with Rabbit IgG TrueBlot®, simply substitute the conventional FITC Anti-rabbit IgG blotting reagent with Fluorescent Rabbit TrueBlot® Antibody Fluorescein and follow the prescribed protocol for sample preparation and immunoblotting. Rabbit IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of rabbit IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Rabbit IgG TrueBlot® with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/Western blot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.

    Purification
    Fluorescent Rabbit TrueBlot® Antibody Fluorescein Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-fluorescein and Anti-Rabbit Serum. Reactivity is observed against native Rabbit IgG by both Western blot and ELISA.
    Labeling Ratio
    3.5
  • Application Notes

    Immunohistochemistry Dilution: User Optimized

    Application Note: Fluorescein Conjugated Antibodies are designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms. Fluorescent Rabbit TrueBlot® Antibody Fluorescein may also be used for detection in immunoassays that do not employ immunoprecipitation. Fluorescent Rabbit TrueBlot® Antibody Fluorescein is provided as a lyophilized powder. To conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). If used conservatively at 2.5 mLs/blot will yield enough reagent for 40 blots. Note that there are three key procedural considerations: 1. Protein A or G should not be used for the immunoprecipitation. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. For immunoprecipitation, Anti-rat IgG beads or Anti-rabbit IgG beads should be used for rat or rabbit immunoprecipitating antibodies, respectively. 2. Immunoprecipitate should be completely reduced. 3. MB-070 Blocking Buffer for Fluorescent Western Blotting should be used as the blocking protein for the immunoblot. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user.

    FLISA Dilution: User Optimized

    Flow Cytometry Dilution: 1:2,000 - 1:10,000

    Western Blot Dilution: 1:1000

    IF Microscopy Dilution: 1:500 - 1:2,500

    Restrictions
    For Research Use only
  • Format
    Lyophilized
    Reconstitution

    Reconstitution Volume: 100 μL

    Reconstitution Buffer: Restore with deionized water (or equivalent)

    Buffer

    Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

    Stabilizer: 10 mg/mL Polyethylene Glycol (PEG-8000)

    Preservative
    Sodium azide
    Precaution of Use
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Storage
    RT,4 °C,-20 °C
    Storage Comment
    Store vial at 4 °C prior to restoration. For extended storage aliquot contents and freeze at -20 °C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4 °C as an undiluted liquid. Dilute only prior to immediate use.
    Expiry Date
    12 months
  • Target Name (Antigen)
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