Human Amylin EIA

$380.00
EIA-AMY-1

RayBio®Human/Mouse/Rat Amylin (IAPP) EIA Kit optimized for serum and cell culture medium. Competition-based ELISA on a 96-well strip plate.

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Antigen Information

Accession Number:
  • P10997
Gene ID:
  • 3375
Gene Symbols:
  • IAPP
Protein Name & Synonyms:
Islet amyloid polypeptide (Amylin) (Diabetes-associated peptide) (DAP) (Insulinoma amyloid peptide)

Assay Format

Detection Range:
0.1-1,000 ng/ml
Sensitivity:
0.62 ng/ml
Specificity:
Cross Reactivity: This EIA kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Nesfatin, Angiotensin II, NPY and APC.
Number of Targets Detected:
1
Species Detected:
  • Human
  • Mouse
  • Rat
Compatible Sample Types:
  • Cell Culture Supernatants
  • Serum
Design Principle:
  • Competition-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Introduction

Amylin, or Islet Amyloid Polypeptide (IAPP), is a 37-residue peptide hormone. It is cosecreted with insulin from the pancreatic beta-cells. Amylin plays a role in glycemic regulation by slowing gastric emptying and promoting satiety, thereby preventing post-prandial spikes in blood glucose levels. IAPP is processed from an 89-residue coding sequence. Proislet Amyloid Polypeptide is produced in the pancreatic beta cells (beta-cells) as a 67 amino acid, 7404 Dalton pro-peptide and undergoes post-translational modifications including protease cleavage to produce amylin.

Insulin and IAPP are regulated by similar factors since they share a common regulatory promoter motif. The IAPP promoter is also activated by stimuli which do not affect insulin, such as tumor necrosis factor alpha and fatty acids. One of the defining features of Type 2 diabetes is insulin resistance. This is a condition wherein the body is unable to utilize insulin effectively, resulting in increased insulin production; since proinsulin and proIAPP are cosecreted, this results in an increase in the production of proIAPP as well.

Amylin's metabolic function is well-characterized as an inhibitor of the appearance of nutrient in the plasma. It thus functions as a synergistic partner to insulin, with which it is cosecreted from pancreatic beta cells in response to meals. The overall effect is to slow the rate of appearance of glucose in the blood after eating; this is accomplished via coordinate slowing down gastric emptying, inhibition of digestive secretion [gastric acid, pancreatic enzymes, and bile ejection, and a resulting reduction in food intake. Appearance of new glucose in the blood is reduced by inhibiting secretion of the gluconeogenic hormone glucagon. These actions, which are mostly carried out via a glucose-sensitive part of the brain stem, the area postrema, may be over-ridden during hypoglycemia. They collectively reduce the total insulin demand.

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

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 Amylin 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.
I used this plate several times along with other similar ELISA kits from different suppliers. I would definitely say that this kit gave me the strongest data.

The instructions were also easy to read and I found the whole procedure to be very simple.

MGH
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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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