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7072
Phototope-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody
WB & IP Reagents
Detection System Kit

Phototope-HRP Western Blot Detection System, Anti-mouse IgG, HRP-linked Antibody #7072

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To Purchase # 7072

Product Usage Information

There are six basic steps in the Western blotting procedures with the Phototope®-HRP Western Blot Detection System.

  1. Polyacrylamide Gel Electrophoresis of Proteins: Separate the protein samples and molecular weight standards by polyacrylamide gel electrophoresis.
  2. Transfer: Transfer the protein to membrane by standard electroblotting.
  3. Block Membrane: Block to saturate nonspecific binding sites on the membrane.
  4. 1° Antibody: Incubate the membrane with the primary antibody.
  5. 2° Antibody: Incubate the membrane with HRP-linked anti-rabbit IgG and HRP-linked anti-biotin antibodies.
  6. Chemiluminescent Detection: Add LumiGLO® reagent and capture the emitted light on X-ray film.

Storage

7076 Anti-mouse IgG, HRP-linked Antibody: Store at -20°C.7727 Biotinylated Protein Ladder Detection Pack: Store at -20°C.7003 20X LumiGLO® Reagent and 20X Peroxide: Store at 4°C.

Product Description

The Phototope-HRP Western Blot Detection System is designed for the chemiluminescent detection of proteins in standard Western blotting applications. Proteins and biotinylated molecular weight markers (provided) are separated by SDS-PAGE and transferred onto membrane. Following incubation with your primary anti-serum, horseradish peroxidase (HRP) linked secondary antibody and HRP-linked anti-biotin antibody are bound and then allowed to react with LumiGLO® reagent. The light emitted by destabilized LumiGLO® reagent is subsequently captured on X-ray film.

Background

Chemiluminescence systems have emerged as the best all-around method for western blot detection. They eliminate the hazards associated with radioactive materials and toxic chromogenic substrates. The speed and sensitivity of these methods are unequalled by traditional alternatives, and because results are generated on film, it is possible to record and store data permanently. Blots detected with chemiluminescent methods are easily stripped for subsequent reprobing with additional antibodies. HRP-conjugated secondary antibodies are utilized in conjunction with specific chemiluminescent substrates to generate the light signal. HRP conjugates have a very high turnover rate, yielding good sensitivity with short reaction times.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.