Fluorescent TrueBlot®: Anti-小鼠 Ig DyLight™ 680

Synonyms: DL680, TrueBlot, DL680 TrueBlot ULTRA, DyLight™ 680 TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, Fluorescent TrueBlot, Ms TrueBlot, IRDye 700, IRDye 680

Background: Mouse IgG TrueBlot® is a unique DyLight™ 680 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 DL680 Anti-mouse IgG blotting reagent with Fluorescent Mouse TrueBlot® Antibody DyLight™ 680 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 blotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.

Immunohistochemistry Dilution: User Optimized

Application Note: Fluorescent Mouse TrueBlot® Antibody DyLight™ 680 may also be used for detection in immunoassays that do not employ immunoprecipitation. Fluorescent Mouse TrueBlot® Antibody DyLight™ 680 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

Reconstitution Volume: 100 μL

Reconstitution Buffer: Restore with deionized water (or equivalent)

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

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