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    Human/Mouse Phospho-Stat 1 (Y701) Cell-Based ELISA Kit

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    2 Citations

    RayBio® Cell-Based Human/Mouse Stat 1 (Tyr701) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.

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    Catalog #:
    CBEL-STAT1
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    Product Description

    Specifications

    Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
    Species Human, Mouse
    Accession Number P42224
    Gene Id 6772
    Gene Symbols STAT1
    Protein Name / Synonyms Signal transducer and activator of transcription 1-alpha/beta (Transcription factor ISGF-3 components p91/p84)
    Quantitative/Semi-Quantitative Semi-Quantitative
    Specificity The antibodies provided in this kit recognizes human and mouse Stat 1 phosphorylated at Tyrosine-701 and total Stat 1 for comparison.
    Compatible Sample Types Adherent Cell Culture
    Solid Support 96-well Microplate
    Method Of Detection Colorimetric
    Design Principle Cell-based
    Research Area Post-Translational Modifications, Phosphorylation, JAK/STAT Signaling
    Estimated Lead Time 1-2 business days
    Shipping Type Blue ice
    Storage -20°C

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    Product Features

    • Site and signal pathway-specific
    • In vitro detection of adherent cell culture
    • No sample lysis is needed
    • Compatible with a standard ELISA plate reader
    • Faster results than with ELISA
    • Adaptable for high-throughput screening and drug discovery

    Application Notes

    Kit Components
    • uncoated 96-well Microplate
    • Wash Buffer A
    • Wash Buffer B
    • Fixing Solution
    • Quenching Buffer
    • Blocking Buffer
    • Anti-phospho antibody
    • Anti-pan antibody
    • HRP-Conjugated Secondary Antibody
    • TMB One-Step Substrate
    • Stop Solution
    Other Materials Required
    • Distilled or deionized water
    • 100 ml and 1 liter graduated cylinders
    • Tubes to prepare sample dilutions
    • Protease and Phosphatase inhibitors
    • Precision pipettes to deliver 2 µl to 1 ml volumes
    • Adjustable 1-25 ml pipettes for reagent preparation
    • Benchtop rocker or shaker
    • Microplate reader capable of measuring absorbance at 450 nm
    Protocol Outline
    1. Seed 10,000-30,000 cells into each well and incubate overnight.
    2. Apply various treatment, inhibitors or activators according to manufacture's instructions.
    3. Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
    4. Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
    5. Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
    6. Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
    7. Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
    8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 µl of Stop Solution to each well.
    11. Read at 450 nm immediately.

    Storage/Stability

    The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

    Citations

    Nakano M., Fujii T., Hashimoto M., et al. Type I interferon induces CX3CL1 (fractalkine) and CCL5 (RANTES) production in human pulmonary vascular endothelial cells. Clin Exp Immunol. 2012 Oct; 170(1): 94-100. doi: 10.1111/j.1365-2249.2012.04638.x
    Species:
    Human
    Sample Type:
    Cell Lysate (Human Pulmonary Artery cells treated with JAK inhibitor)
    View Publication
    Valerio, Michael, and Atif B. Awad. "?-Sitosterol down-regulates some pro-inflammatory signal transduction pathways by increasing the activity of tyrosine phosphatase SHP-1 in J774A. 1 murine macrophages." International immunopharmacology 11.8 (2011): 1012-1017.
    Species:
    Human
    Sample Type:
    Cell Lysate
    View Publication

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