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Phosphorylation involves the addition of phosphate groups to proteins, most commonly at a serine (S), threonine (T) or tyrosine (Y) residue by a kinase and is essential for pathway activation in cellular regulation, cell signaling and growth. Our portfolio offers a large and diverse set of phospho specific antibodies and reagents that detect phosphorylated targets with great precision. These phospho antibodies and phospho reagents can be used in western blot, flow cytometry, immunohistochemistry and immunofluorescence microscopy. Additionally, Rockland phospho antibodies provide accuracy and dependability that clients within the life sciences have come to expect.
Protein phosphorylation plays a significant role in a wide range of cellular processes including cell growth and proliferation, metabolism, physiological regulation and cell signaling. Activated phospho proteins and phospho reagents usually trigger a cascade of reactions and interactions which lead to cellular activation, motility, proliferation and metabolic modifications. Due to the complex roles that phosphorylated proteins can play, accurately and precisely quantifying phosphorylated proteins at the cellular level is critical to deciphering and understanding cellular functions. At Rockland, we have developed a vast and diverse portfolio of phospho specific antibodies and phospho reagents that allow scientists to track and quantify phospho proteins using a wide range of techniques such as phospho immunoprecipitation western blot, flow cytometry, immunohistochemistry and immunofluorescence.
Oxidative phosphorylation is a process that can provide a number of fascinating results and insightful realizations within the life sciences. With more than thirty years of experience, Rockland’s portfolio has grown to include an impressive collection of phospho specific antibodies and reagents. Our philosophy is to constantly meet new challenges by continuously expanding our collection of phospho specific antibodies for applications in neuroscience, cellular growth and metabolism, cancer research, cell signaling, epigenetics, apoptosis, autophagy and responses to the extra-cellular environment. Our phospho antibodies are unparalleled in quality, precision, and accuracy.
Rockland offers a wide range of monoclonal and polyclonal antibodies specific for activated Akt at positions pT308 and pS473.
The Akt family of serine-threonine kinases consists of three members: Akt-1, Akt-2 and Akt-3. All Akt family members share a common structure with an N-terminal plekstrin domain, a central kinase domain and C-terminal hydrophobic domain. The Akt family members are not redundant and regulate a broad spectrum of cellular functions and cell signaling, however to be fully activated, Akt requires phosphorylation at two specific sites: threonine 308 and serine 473. Upon activation, Akt moves from the membrane to the cytoplasm and nucleus where it phosphorylates its target.
Rockland offers a wide variety of primary and secondary phospho specific antibodies validated by a variety of immunoassays including western blot protocol, immunohistochemistry and flow cytometry. Antibodies recognize phosphorylated target proteins involved in the apoptotic pathway, the cell signaling cycle, stem cell development, cancer and other pathways of critical interest to life science researchers.
Rockland's phospho specific conjugated antibodies are useful because they eliminate the need for a secondary antibody. By conjugating the primary antibodies, you can set up a direct assay and avoid potential cross-reactivity of the secondary antibodies. The direct phospho assay uses the method of directly labeling the monoclonal antibody itself. The antibody is conjugated with a colorimetric, chemiluminescent or fluorescent label. Since the secondary antibody step is omitted, the direct assay is fast and avoids potential problems of cross-reactivity of the phospho specific antibody with components in the antigen sample.
Rockland's TrueBlot® reagents preferentially detects the native disulfide form of mouse IgG or rabbit IgG, making it ideal for IP western blot protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot® reduces interference by the ~55 kDa heavy and ~23 kDa light chains of the immunoprecipitating antibody in IP/immunoblotting applications. TrueBlot® is also ideal for studying post-translational protein modifications, including phosphorylation, or protein-protein interactions.
TrueBlot® enables researchers to generate publication-quality IP western blot data. Simply substitute your conventional HRP blotting reagent with Goat TrueBlot®, Mouse TrueBlot®, Rabbit TrueBlot® or Sheep TrueBlot®. TrueBlot® is a horseradish peroxidase conjugated immunoblotting detection reagent, which enables unhindered detection of blotted target bands. This results in unparalleled clarity in results and high quality imaging suitable for reproduction.
Rockland can develop monoclonal or polyclonal custom phospho specific antibodies to the phosphorylation sites of interest. Critical to the development of a custom phosphorylated antibody is the design of the immunizing peptide. The phosphorylated antibody is then affinity purified using both positive and negative adsorption methodologies to optimize the desired reactivity. Phospho ELISA testing against both phosphorylated and non-phosphorylated peptides confirms the success of the antibody purification.
The Mouse TrueBlot® Western Blot Kit contains the critical supporting reagents, buffers, and substrates for Phospho immunoprecipitation and IP Western blotting of samples using TrueBlot second step immunoblotting reagents in conjunction with your own primary IP antibody and primary (Mouse IgG) Western blotting antibody. TrueBlot technology enables unhindered detection of protein bands of interest which would otherwise be obscured by the presence of reduced and denatured heavy and light chain immunoglobulin in the blot (as detected by the conventional immunoblotting HRP anti-mouse IgG reagent). Mouse IgG TrueBlot® ULTRA is the unique horseradish peroxidase conjugated anti-mouse IgG immunoblotting second step reagent which enables detection of Phospho immunoblotted target protein bands, without hindrance by interfering immunoglobulin heavy and light chains from your IP antibody. Use it in place of your usual HRP anti-mouse IgG immunoblotting second step reagent. It is easy to generate publication-quality Phospho IP Western Blot 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 Phospho IP western blotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.
Rockland Immunochemicals Inc.Gilbertsville, PA 19525E-mail: email@example.comPhone: 800.656.7625