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Fluorescent TrueBlot®: Anti-Mouse Ig Fluorescein

Catalog No. ABIN6698838
  • Target See all Ig products
    Ig
    Reactivity
    Mouse
    Host
    Rat
    Conjugate
    FITC
    Application
    Immunohistochemistry (IHC), Western Blotting (WB), FLISA, Flow Cytometry (FACS), Fluorescence Microscopy (FM)
    Brand
    TrueBlot®
    Characteristics

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

    Background: Mouse IgG TrueBlot® is a unique fluorescein conjugated Anti-mouse IgG immunoblotting (second step) reagent. Mouse 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 Mouse IgG TrueBlot®, simply substitute the conventional FITC Anti-mouse IgG blotting reagent with Fluorescent Mouse TrueBlot® Antibody Fluorescein and follow the prescribed protocol for sample preparation and immunoblotting. Mouse IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of mouse IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Mouse 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 Mouse 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-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA.
    Labeling Ratio
    3.6
  • Application Notes

    Immunohistochemistry Dilution: User Optimized

    Application Note: Fluorescent Mouse TrueBlot® Antibody Fluorescein may also be used for detection in immunoassays that do not employ immunoprecipitation. 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 Mouse 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 2.5mls/blot will yield enough reagent for 40 blots. Note that there are three key procedural considerations: 1. Protein A or G beads may be used with the mouse, goat and sheep TrueBlot secondaries, but not with the rabbit TrueBlot secondary. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. 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. Note: To achieve best results when detecting mouse IgG1 subtypes, we recommend performing a dot blot or titration to determine the optimal dilution factor for your desired application. 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: User Optimized

    Western Blot Dilution: 1:1000

    IF Microscopy Dilution: 1:200

    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)

    0.01 % (w/v) Sodium Azide
    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
    Ig
    Abstract
    Ig Products
    Synonyms
    ATPIG Accessory Reagents, ATPIQ Accessory Reagents, A330005H02Rik Accessory Reagents, AI315324 Accessory Reagents, Ig Accessory Reagents, ATPase phospholipid transporting 11C Accessory Reagents, ATPase, class VI, type 11C Accessory Reagents, ATP11C Accessory Reagents, Atp11c Accessory Reagents
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