MSH alpha ELISA Kit

Catalog No. ABIN8070080

Human, Mouse and Rat MSH alpha ELISA Kit, Colorimetric assay for quantification of Human, Mouse and Rat MSH alpha.

Quick Overview for MSH alpha ELISA Kit (ABIN8070080)

Target: MSH alpha

Reactivity: Human, Mouse, Rat

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 0.98 pg/mL - 1000 pg/mL

Application: ELISA

Sample Type: Plasma, Serum

Product Details

Minimum Detection Limit: 0.98 pg/mL

Purpose: 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.

Analytical Method: Quantitative

Specificity: This EIA kit is designed to detect human mouse, and rat Alpha-Melanocytestimulating hormone.

Sensitivity: 20.5 pg/mL

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

Material not included:

• 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 450 nm
• SigmaPlot software (or other software that can perform four-parameter logistic regression models)

Application Details

Application Notes: Suggested Plasma Dilution: Human: 4X / Mouse: 8X / Rat: 8X

Sample Volume: 100 μL

Plate: Pre-coated

Protocol:

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.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: 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°C to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.

Target Details

Target: MSH alpha

Alternative Name: alpha-MSH

Background:

Gene Symbols: STAMBP, AMSH

Background: 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.

Protein Name (Synonyms): alpha-melanocyte stimulating hormone (a-MSH/ aMSH)

Gene ID: 10617

UniProt: O95630

Pathways: Metabolism of Steroid Hormones and Vitamin D, Peptide Hormone Metabolism, Hormone Activity