Insulin ELISA Kit

Catalog No. ABIN8069803

The Human Insulin ELISA Kit (ABIN8069803) is a Colorimetric ELISA Kit designed to quantify Human Insulin.

Quick Overview for Insulin ELISA Kit (ABIN8069803)

Target: Insulin (INS)

Binding Specificity: total, ubiquitinated

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details Insulin ELISA Kit

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

Analytical Method: Quantitative

Components:

Size / Qty
• Microplate Coated With Anti-Pan-Insulin Antibody
• Anti-Ubiquitination Detection Antibody
• Anti-Insulin Detection Antibody
• HRP-Conjugated Streptavidin
• 200 μL
• Wash Buffer
• Assay Diluent
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• 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 sample dilutions
• Distilled or deionized water
• Shaker

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed.
2. Add 100 μL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

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

Target Details for Insulin

Target: Insulin (INS)

Alternative Name: Insulin

Background:

Gene Symbols: INS

Background: The Ubiquitinated-Insulin and Total-Insulin ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human ubiquitinated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan Insulin antibody has been coated onto a 96-well plate. 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 biotinylated anti-ubiquitination antibody is used to detect ubiquitinated Insulin or biotinylated anti-Insulin antibody is used to detect total Insulin . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of ubiquitinated Insulin or total 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

Gene ID: 3630

UniProt: P01308

Pathways: NF-kappaB Signaling, RTK Signaling, 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