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    Alpha-MSH ELISA Kit

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    100 % of 100
    1  Review (include details such as sample type used, experimental conditions, etc.)
    The RayBio® Alpha-MSH Enzyme Immunoassay (EIA) Kit is an in vitro quantitative assay for detecting human, mouse, or rat melanocyte-stimulating hormone alpha based on the competitive ELISA principle. The immunoassy is optimized for serum, plasma and cell culture media.
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    EIA-aMSH
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    Product Description

    Specifications

    Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
    Species Human, Mouse, Rat
    Accession Number P01189#PRO_0000024970
    Protein Name / Synonyms alpha-melanocyte stimulating hormone (a-MSH/ aMSH)
    Quantitative/Semi-Quantitative Quantitative
    Specificity This EIA kit is designed to detect human mouse, and rat Alpha-Melanocytestimulating hormone.
    Compatible Sample Types Cell Culture Supernatants, Serum
    Solid Support 96-well Microplate
    Method Of Detection Colorimetric
    Design Principle Competition-based
    Sensitivity 20.5 pg/ml
    Detection Range 0.98-1,000 pg/ml
    Recommended Dilution (Serum/Plasma) Human: 4X / Mouse: 8X / Rat: 8X
    Estimated Lead Time 1-2 business days
    Shipping Type Blue ice
    Storage -20°C

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    Introduction

    Alpha-Melanocyte-stimulating hormone (alpha-MSH), a proteolyic cleavage product from ACTH (1-13), is an endogenous peptide hormone and neuropeptide of the melanocortin family. It is a non-selective full agonist of the melanocortin receptors MC1, MC3, MC4, and MC5, but not MC2. Alpha-MSH is important for pigmentation primarily of the hair and skin. It also plays a role in feeding behavior, energy homeostasis, sexual activity, and protection against ischemia and reperfusion injury.

    In this assay, a biotinylated Alpha-MSH peptide is spiked into the samples and standards. The samples and standards are then added to the plate, where the biotinylated Alpha-MSH peptide competes with endogenous (unlabeled) Alpha-MSH for binding to the anti-Alpha-MSH antibody. After a wash step, any bound biotinylated Alpha-MSH then interacts with horseradish peroxidase (HRP)- streptavidin, which catalyzes a color development reaction. The intensity of the colorimetric signal is directly proportional to the amount of captured biotinylated Alpha-MSH peptide and inversely proportional to the amount of endogenous Alpha- MSH in the standard or samples. A standard curve of known concentration of Alpha-MSH peptide can be established and the concentration of Alpha-MSH peptide in the samples can be calculated accordingly.

    Product Features

    • Strip plates and additional reagents allow for use in multiple experiments
    • Quantitative protein detection
    • Establishes normal range
    • The best products for confirmation of antibody array data

    Typical Data

    Standard Curve
    Human Alpha-MSH EIA Standard Curve

    Application Notes

    Kit Components
    • Pre-Coated 96-well Strip Microplate
    • Wash Buffer
    • Standard Peptide
    • Assay Diluent(s)
    • Biotinylated Peptide
    • HRP-Streptavidin
    • TMB One-Step Substrate
    • Stop Solution
    • Assay Diagram
    • Positive Control Sample
    • Capture Antibody
    • User Manual
    Other Materials Required
    • Distilled or deionized water
    • Precision pipettes to deliver 2 µl to 1 ml volumes
    • Adjustable 1-25 ml pipettes for reagent preparation
    • 100 ml and 1 liter graduated cylinders
    • Tubes to prepare standard and sample dilutions
    • Orbital shaker
    • Aluminum foil
    • Saran Wrap
    • Absorbent paper
    • Microplate reader capable of measuring absorbance at 450nm
    • SigmaPlot software (or other software that can perform four-parameter logistic regression models)
    Protocol Outline
    1. Prepare all reagents, samples and standards as instructed.
    2. Add 100 µl detection antibody to each well.
    3. Incubate 1.5 h at RT or O/N at 4°C.
    4. Add 100 µl standard or sample to each well.
    5. Incubate 2.5 h at RT.
    6. Add 100 µl prepared streptavidin solution.
    7. Incubate 45 min at RT.
    8. Add 100 µl TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 µl Stop Solution to each well.
    11. Read plate at 450 nm immediately.

    Storage/Stability

    Standard, Biotinylated Alpha-MSH peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.

    Reviews

    Write a customer review
    1. Perfect result was observed
      Rating
      100%
      This Human Alpha-MSH EIA Kit works well for human serum samples. The kit is easy to use and the sensitivity is high.

      Review by

      from The University of Texas MD Anderson Cancer Center,

      on

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