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525282 PhosphoDetect™ Phosphoserine Detection Kit

525282
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      Description
      OverviewA set of four different monoclonal antibodies specific for serine phosphorylation sites. Recognition is dependent on phosphorylation and the surrounding amino acid motif. Recognizes serine phosphorylated proteins in Arabidopsis, bacteria, chicken, human, maize, mouse, rat, tobacco, Xenopus, yeast, and zebrafish. Phosphorylation patterns in cell extracts may differ when probed with different antibodies.
      Catalogue Number525282
      Brand Family Calbiochem®
      Application Data

      Detection of phosphoserine-containing antibodies by immunoblotting. Samples: Whole cell lysate from A431 cells treated with pervanadate (all lates). Primary antibodies: PhosphoDetect™ Anti-Phosphoserine Mouse mAb (16B4) (Cat. No. 525280) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (1C8) (Cat. No. 525281) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A3) (Cat. No. 525283) (1 µg/ml), and PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A9) (Cat. No. 525284) (1 µg/ml). Detection: chemiluminescence.

      Detection of phosphoserine-containing antibodies by immunoblotting. Samples: Extract from rabbit muscle*. Primary antibodies: PhosphoDetect™ Anti-Phosphoserine Mouse mAb (16B4) (Cat. No. 525280) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (1C8) (Cat. No. 525281) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A3) (Cat. No. 525283) (1 µg/ml), and PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A9) (Cat. No. 525284) (1 µg/ml). Detection: chemiluminescence. *Positive control supplied with the antibody.
      References
      Product Information
      FormLyophilized
      Formulation25 µg each antibody lyophilized from 2X PBS, PEG, sucrose and 200 µl control phosphoproteins lyophilized 20 mM Na₂HPO₄, 0.1% SDS.
      Kit contains25 μg of each of the Phosphoserine Antibodies including clones 1C8, 4A3, 4A9, and 16B4, Rabbit Muscle Positive Control, and a data sheet.
      Positive controlPhosphoproteins purified from rabbit muscle (included)
      Preservative≤ 0.1% sodium azide (antibodies only)
      Quality LevelMQ100
      Applications
      Key Applications Enzyme-Linked Immunosorbent Assay
      Immunoblotting (Western Blotting)
      Immunoprecipitation
      Application NotesImmunoblotting (see characteristics table)
      Immunoprecipitation (see characteristics table)
      ELISA (see characteristics table)
      Application CommentsDO NOT use milk or casein for membrane blocking or antibody dilutions; use BSA. For immunoblotting with the positive control, use 20 µl/lane (minigel) for colorimetric detection of 5 µl/lane for chemiluminescent detection. The phosphorylation patterns in a given cell lysate may differ when probed with individual antibodies due to sequence specificity. Antibodies should be titrated for optimal results in individual systems.
      Biological Information
      Immunogenphosphoserine-containing peptides or phosphoserine conjugated to KLH
      HostMouse
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Standard Handling
      Storage -20°C
      Do not freeze Ok to freeze
      Special InstructionsReconstitute each antibody with 1 ml H₂O (15 min, room temperature). Following reconstitution, aliquot and freeze (-70°C). Stock solutions are stable for up to 1 year at -70°C. Thaw aliquots at 37°C prior to use. Reconstitute the positive control with 200 µl H₂O. After complete solubilization, add 200 µl SDS-PAGE sample buffer and incubate for 5 min at 90°C. Following incubation, aliquot and freeze (-20°C). Avoid freeze/thaw cycles of all solutions.
      Packaging Information
      Transport Information
      Supplemental Information
      Kit contains25 μg of each of the Phosphoserine Antibodies including clones 1C8, 4A3, 4A9, and 16B4, Rabbit Muscle Positive Control, and a data sheet.
      Specifications

      Documentation

      PhosphoDetect™ Phosphoserine Detection Kit Certificates of Analysis

      TitleLot Number
      525282
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision20-April-2009 JSW
      ApplicationImmunoblotting (see characteristics table)
      Immunoprecipitation (see characteristics table)
      ELISA (see characteristics table)
      Application Data

      Detection of phosphoserine-containing antibodies by immunoblotting. Samples: Whole cell lysate from A431 cells treated with pervanadate (all lates). Primary antibodies: PhosphoDetect™ Anti-Phosphoserine Mouse mAb (16B4) (Cat. No. 525280) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (1C8) (Cat. No. 525281) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A3) (Cat. No. 525283) (1 µg/ml), and PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A9) (Cat. No. 525284) (1 µg/ml). Detection: chemiluminescence.

      Detection of phosphoserine-containing antibodies by immunoblotting. Samples: Extract from rabbit muscle*. Primary antibodies: PhosphoDetect™ Anti-Phosphoserine Mouse mAb (16B4) (Cat. No. 525280) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (1C8) (Cat. No. 525281) (1 µg/ml), PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A3) (Cat. No. 525283) (1 µg/ml), and PhosphoDetect™ Anti-Phosphoserine Mouse mAb (4A9) (Cat. No. 525284) (1 µg/ml). Detection: chemiluminescence. *Positive control supplied with the antibody.
      DescriptionA set of 4 mouse monoclonal antibodies purified from serum-free culture supernatant by thiophilic adsorption and size exclusion chromatography. All 4 antibodies recognize phosphoserine. Supplied with a positive control consisting of phosphoproteins purified from rabbit muscle lysate by Fe3+-IDA affinity chromatography.
      BackgroundProtein phosphorylation is the principal indicator of the activation of signaling pathways. The binding of a growth factor to its specific receptor catalyzes a complex cascade of intracellular signaling events, characterized by altering the phosphorylation of tyrosine, serine, or threonine residues of target proteins. The phosphorylation state is tightly regulated by the actions of protein kinases, which transfer a phosphate group from donor substrates such as ATP or GTP to serine, threonine, or tyrosine residues, and protein phosphatases, which dephosphorylate phosphorylated proteins, thereby restoring the particular phosphorylation system to its basal stage. Most kinases are either serine/threonine-specific or tyrosine-specific, while some kinases can phosphorylate all 3 types of hydroxy-bearing amino acids. Radioactive kinase assays utilizing 32P-γ-ATP as the donor substrate have been frequently employed to assess kinase activity, since the method is sensitive, convenient, and easily quantified. Recently, however, the generation of phosphoprotein-specific antibodies has advanced the development of non-radioactive kinase activity detection methods to comparable sensitivity as radioactive methods. Antibodies specific for phosphorylated epitopes recognize the phosphorylated amino acid in the context of the surrounding amino acid sequence. Recognition, therefore, depends on 2 criteria: 1) phosphorylation and 2) the surrounding amino acid motif. If one of the two criteria is not fulfilled, the antibody will not detect the phosphorylation site. Since the surrounding amino acid sequence varies between phosphorylation sites, certain proteins, though phosphorylated, may not be detected by a given antibody.
      HostMouse
      Immunogenphosphoserine-containing peptides or phosphoserine conjugated to KLH
      Positive controlPhosphoproteins purified from rabbit muscle (included)
      FormLyophilized
      Formulation25 µg each antibody lyophilized from 2X PBS, PEG, sucrose and 200 µl control phosphoproteins lyophilized 20 mM Na₂HPO₄, 0.1% SDS.
      Preservative≤ 0.1% sodium azide (antibodies only)
      CommentsDO NOT use milk or casein for membrane blocking or antibody dilutions; use BSA. For immunoblotting with the positive control, use 20 µl/lane (minigel) for colorimetric detection of 5 µl/lane for chemiluminescent detection. The phosphorylation patterns in a given cell lysate may differ when probed with individual antibodies due to sequence specificity. Antibodies should be titrated for optimal results in individual systems.
      Storage -20°C
      Do Not Freeze Ok to freeze
      Special InstructionsReconstitute each antibody with 1 ml H₂O (15 min, room temperature). Following reconstitution, aliquot and freeze (-70°C). Stock solutions are stable for up to 1 year at -70°C. Thaw aliquots at 37°C prior to use. Reconstitute the positive control with 200 µl H₂O. After complete solubilization, add 200 µl SDS-PAGE sample buffer and incubate for 5 min at 90°C. Following incubation, aliquot and freeze (-20°C). Avoid freeze/thaw cycles of all solutions.
      Toxicity Standard Handling

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