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Datasheet

Goat TrueBlot® Western Blot Kit

Mouse Monoclonal eB270

88-8884-31
1 Kit
Goat
Mouse
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Description

Background

The Goat TrueBlot® Western Blot Kit contains the critical supporting reagents, buffers, and substrates for immunoprecipitation and Western blot of samples using TrueBlot monoclonal secondary antibody in conjunction with your own primary IP antibody and primary (Goat IgG) Western blot antibody (see application note for kit components). Goat IgG TrueBlot® is the unique horseradish peroxidase conjugated Anti-Goat IgG monoclonal secondary antibody which enables detection of immunoblotted target protein bands, without hindrance by interfering immunoglobulin heavy and light chains from your IP antibody. Use it in place of your conventional HRP Anti-Goat IgG immunoblotting secondary antibody. It is easy to generate publication‐quality IP/WB data with Goat IgG TrueBlot. Goat IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of goat IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Goat 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/immunoblotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions. Goat IgG TrueBlot® may also be used for detection in immunoblotting assays that do not employ immunoprecipitation.

Synonyms

Anti-Goat IgG HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, TrueBlot for chemiluminescent western blotting

Application Note

Goat IgG TrueBlot® is provided as 1000X solution. 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 20 blots. Goat IgG TrueBlot® HRP-conjugated monoclonal secondary antibody reacting with goat IgGs for optimal signal detection in immunoprecipitation/immunoblotting experiments. 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. 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. BLOTTO/Milk should be used as the blocking protein for the immunoblot. A sample of Anti-Alpha-Tubulin (MOUSE) Monoclonal Antibody Cat #(200-301-880S), has been provided to be used as a loading control at the user’s discretion. Dilutions are to be optimized by the user. Goat TrueBlot Western Blot Kit Components: 1. Goat IgG TrueBlot: 50 μL. 18-8814-31; 2. TrueBlot Enhancer Solution: 25 mL; 3. TrueBlot Blocker: 10 g; 4. TrueBlot Assay Buffer: 30 mL, 20X; 5. TrueBlot Substrate A: 12.5 mL; 6. TrueBlot Substrate B: 12.5 mL; 7. Western Blot Incubation Tray. 8. Anti-Alpha-Tubulin (MOUSE) Monoclonal Antibody - 200-301-880S.

Purity/Specificity

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

Storage Condition

Store Kit at 2-8 °C, except Goat TrueBlot® which should be stored at -20 °C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.

Disclaimer Note-General

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.

Goat TrueBlot® Western Blot Kit - 88-8884-31
Goat
Goat
Mouse
Peroxidase (Horseradish) ULTRA
Monoclonal
eB270
IgG
Immunoprecipitation Kit
Liquid (sterile filtered)
Wet Ice
Expiration date is six (6) months from date of opening.
0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol

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