TrueBlot® Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag
Mouse TrueBlot® IP / Western Blot
Anti-6X His Epitope Tag Monoclonal Antibody - Western Blot
Mouse TrueBlot® Western Blot Kit
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Datasheet

TrueBlot® Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag

Monoclonal 33D10.D2.G8

88-8887-382
1 Kit
IP, WB
6X His-Tag, Mouse
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$459.00 /Per Item
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Description

Background

TrueBlot® Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag allows for the immunoprecipitation and detection of 6X HIS-tagged recombinant proteins present in cell lysates provided by the user. After protein separation by SDS-PAGE and transfer, the membrane is probed with monoclonal Anti-6X HIS. Detection of the membrane bound antibody-antigen complex is achieved by the addition of Mouse IgG TrueBlot® ULTRA HRP. The TrueBlot® Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag contains the critical supporting reagents, buffers, and substrates for immunoprecipitation and Western blotting of samples containing a 6X HIS Epitope Tag using TrueBlot monoclonal secondary antibody in conjunction with Rockland's Anti-6X HIS (MOUSE) Monoclonal Antibody. Mouse IgG TrueBlot® ULTRA is the unique horseradish peroxidase conjugated Anti-Mouse IgG monoclonal secondary antibody which enables detection of immunoblotted target protein bands, without hindrance from interfering immunoglobulin heavy and light chains from your IP antibody. Use it in place of your conventional HRP Anti-Mouse IgG immunoblotting secondary antibody. It is easy to generate publication-quality IP/WB data with Mouse IgG TrueBlot® ULTRA. Mouse IgG TrueBlot ULTRA 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 ULTRA 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.

Application Note

Mouse IgG TrueBlot® ULTRA 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). Mouse IgG TrueBlot® ULTRA is an HRP-conjugated monoclonal secondary antibody reacting with mouse IgGs for optimal signal detection in immunoprecipitation/immunoblotting experiments. 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. TrueBlot® IP and WB Kit for 6X HIS Epitope Tag Components: 1. Mouse IgG TrueBlot ULTRA: 50 μL 18-8817-31; 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. Anti-Mouse Ig IP Beads: 2.5 mL Binds 0.4 mg Ig/mL beads; 8. Anti-6X HIS (MOUSE) Monoclonal Antibody: 100 µg; 9. Western Blot Incubation Tray. Special Notes: Upon initial use of the IP beads, we recommend that the vial be inverted several times to get the beads into suspension. We recommend using a large bore pipet to pipet up the liquid for use. For storage of the opened vial of beads, we recommend that the vial cap be sealed with parafilm to help prevent evaporation of the buffer. 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

This TrueBlot® Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag allows for the detection of 6X HIS-tagged recombinant protein present in cell lysates provided by the user. Mouse TrueBlot® ULTRA 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-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA.

TrueBlot® Immunoprecipitation and Western Blot Kit for 6X HIS Epitope Tag - 88-8887-382
HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, 6X HIS Western Blot, Western Blotting Kit, Chemiluminescent 6X HIS Kit, Immunoblotting Kit, HRP Anti 6X HIS, 6X HIS Epitope Tag, 6X HIS, IP, H-H-H-H-H-H Tag, Epitope Tag
IP, WB
Mouse
6X His-Tag, Mouse
6X HIS
Peroxidase (Horseradish) ULTRA
Monoclonal
33D10.D2.G8
IgG
Immunoprecipitation Kit
Liquid (sterile filtered)
0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Wash buffers MUST NOT contain SODIUM AZIDE or other inhibitors of peroxidase activity!
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol
Wet Ice
Store Kit at 2-8 °C, except Mouse TrueBlot® ULTRA and Anti-6X HIS (MOUSE) Monoclonal Antibody, which should be stored at -20 °C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.
Expiration date is six (6) months 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.