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    Phospho-Stat 3 (Y705) ELISA Kit

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    5 Citations

    RayBio® Human/Mouse/Rat Phospho-Stat 3 (Y705) ELISA Kit. This assay semi-quantitatively measures phosphorylated Stat 3 (Tyr705) in lysate samples.

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    Catalog #:
    PEL-Stat3-Y705
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    Product Description

    Specifications

    Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
    Species Human, Mouse, Rat
    Accession Number P40763
    Gene Id 6774
    Gene Symbols STAT3|APRF
    Protein Name / Synonyms Signal transducer and activator of transcription 3 (Acute-phase response factor)
    Quantitative/Semi-Quantitative Semi-Quantitative
    Specificity The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Stat3 (pTyr705).
    Compatible Sample Types Tissue Lysates, Cell Lysates
    Solid Support 96-well Microplate
    Method Of Detection Colorimetric
    Design Principle Sandwich-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

    • Rapidly measure phosphorylated protein in lysates
    • Screen numerous different cell lysates without performing a Western Blot analysis
    • Minimal hands-on time, convenient, and non-radioactive material

    Application Notes

    Kit Components
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Anti-Phospho Antibody
    • HRP-Conjugated Secondary Antibody
    • Assay Diluent
    • TMB One-Step Substrate
    • Stop Solution
    • Lysis Buffer
    • Positive Control Sample
    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. Prepare all reagents and samples as instructed in the manual.
    2. Add 100 µl of sample or positive control to each well.
    3. Incubate 2.5 h at RT or O/N at 4 °C.
    4. Add 100 µl of prepared primary antibody to each well.
    5. Incubate 1 h at RT.
    6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.
    7. Incubate 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

    Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

    Citations

    Przybyla, Laralynne M., et al. "Matrix Remodeling Maintains Embryonic Stem Cell Self?Renewal by Activating Stat3." Stem cells 31.6 (2013): 1097-1106.
    Species:
    Human
    Sample Type:
    Conditioned Media (embryonic stem cells)
    View Publication
    Di Grazia A., Cappiello F., Imanishi A., et al. The Frog Skin-Derived Antimicrobial Peptide Esculentin-1a(1-21)NH2 Promotes the Migration of Human HaCaT Keratinocytes in an EGF Receptor-Dependent Manner: A Novel Promoter of Human Skin Wound Healing? PLoS One. 2015 Jun 12;10(6):e0128663. doi: 10.1371/journal.pone.0128663
    Species:
    Human
    Sample Type:
    Cell Lysate (HaCaT cells (human immortalized keratinocyte cells)
    View Publication
    Sharma S., et al. IL-10 directly protects cortical neurons by activating PI-3 kinase and STAT-3 pathways. Brain Research Volume 1373, 10 February 2011, Pages 189-194.
    Species:
    Rat
    Sample Type:
    Cell Lysate (Primary Rat neuronal cells)
    View Publication
    Iman O. Sherif & Nora H. Al-Shaalan (2020): Hepatoprotective effect of Ginkgo biloba extract against methotrexate-induced hepatotoxicity via targeting STAT3/miRNA-21 axis, Drug and Chemical Toxicology
    Species:
    Rat
    Sample Type:
    Tissue Lysate (liver)
    View Publication
    Mohamed, Y.S., Abdelsalam, R.M., Attia, A.S. et al. Regulation of liver regeneration by prostaglandin E2 and thromboxane A2 following partial hepatectomy in rats. Naunyn-Schmiedeberg's Arch Pharmacol 393, 1437-1446 (2020). https://doi.org/10.1007/s00210-0
    Species:
    Rat
    Sample Type:
    Tissue Lysate (Liver)
    View Publication

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