Insulin Kit ELISA

N° du produit ABIN8069778

Ce kit ELISA Colorimetric est destiné à la mesure quantitative de Humain Insulin.

Aperçu rapide pour Insulin Kit ELISA (ABIN8069778)

Antigène: Insulin (INS)

Épitope: Oxidized Form, total

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Type d'échantillon: Cell Lysate, Tissue Lysate

Détail du produit Insulin Kit ELISA

Fonction: Measures oxidated Insulin (INS) and total Insulin in human cell and tissue lysates, enabling precise assessment of Insulin oxidation by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.

Analytical Method: Quantitative

Ingrédients:

Size / Qty
• Microplate Coated With Anti-Pan-Insulin Antibody
• Positive Control
• HRP-Conjugated Streptavidin
• 200 μL
• Wash Buffer
• Assay Diluent
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution
• Oxidation Buffer A
• Oxidation Buffer B
• Oxidation Labeling Reagent
• Oxidation Stabilizing Reagent

Matériel non inclus:

• 20 % (v/v) trichloroacetic acid (TCA) solution, ice-cold
• 1:1 (v/v) ethanol:ethyl acetate
• Benchtop centrifuge and microcentrifuge (4 °C)
• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• 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
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare positive control or sample dilutions
• Distilled or deionized water
• Shaker

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and Positive Control as instructed.
2. Label samples with Oxidation Labeling Reagent
3. Add Oxidation Stabilizing Reagent to samples.
4. Precipitate proteins with ice-cold 20 % TCA.
5. Wash protein pellet twice with 1:1 (v/v) ethanol:ethyl acetate.
6. Resuspend protein pellets in Oxidation Buffer B.
7. Add 100 μL sample and Positive Control to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
8. Add 100 μL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
9. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
10. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Stockage

Stock: -20 °C

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

Détail du antigène

Antigène: Insulin (INS)

Autre désignation: Insulin

Sujet:

Gene Symbols: INS

Background: The Oxidated Insulin ELISA Kit uses the "biotin probes" method to biotin label carbonyl groups of oxidized proteins. In this method, the biotin probes first react with carbonyl groups of oxidized proteins to form unstable Schiff bases, which are then further reduced to more stable amines. The biotinylated proteins can then be detected by ELISA. An anti-pan Insulin antibody has been coated onto a 96-well plate. Biotinylated samples are pipetted into the wells, and Insulin present in a sample is bound to the wells by the immobilized antibody. The wells are washed, and HRP-conjugated streptavidin is used to detect oxidated Insulin. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of oxidated Insulin bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): Insulin

ID gène: 3630

UniProt: P01308

Pathways: Signalisation NF-kappaB, Signalisation RTK, Positive Regulation of Peptide Hormone Secretion, Peptide Hormone Metabolism, Hormone Activity, Carbohydrate Homeostasis, ER-Nucleus Signaling, Regulation of Carbohydrate Metabolic Process, Feeding Behaviour, Autophagy, Negative Regulation of intrinsic apoptotic Signaling, Brown Fat Cell Differentiation, Positive Regulation of fat Cell Differentiation