Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680
Rabbit TrueBlot® IP / Western Blot
Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight 680 Conjugated - Western Blot
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Datasheet

Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680

Mouse Monoclonal eB182

18-4416-32
100 µL
IP, WB
Rabbit
Mouse
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Description

Background

Rabbit IgG TrueBlot® is a unique DyLight™ 680 conjugated Anti-rabbit IgG monoclonal secondary antibody. 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 DL680 Anti-rabbit IgG blotting reagent with Fluorescent Rabbit TrueBlot® Antibody DyLight™ 680 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.

Application Note

Fluorescent Rabbit TrueBlot® Antibody DyLight™ 680 has been tested in western blot and immunoprecipitation and may also be used for detection in immunoassays that do not employ immunoprecipitation. Fluorescent Rabbit 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 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.

Purity/Specificity

Fluorescent Rabbit TrueBlot® Antibody DyLight™ 680 Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against Anti-Rabbit Serum. Reactivity is observed against native Rabbit IgG by both Western blot and ELISA.

Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680 - 18-4416-32
Anti-Rabbit IgG DL680, TrueBlot, DL680 TrueBlot ULTRA, DyLight™ 680 TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, Fluorescent TrueBlot, Rb TrueBlot
IP, WB
Rabbit
Mouse
DyLight™ 680
Monoclonal
eB182
IgG
2.3 moles DL680 per mole of IgG
Lyophilized
1.0 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
10 mg/ml Polyethylene Glycol (PEG-8000)
100 µL
Restore with deionized water (or equivalent)
Ambient
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.
Expiration date is one (1) year from date of opening.
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This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.