GCK ELISA Kit

Catalog No. ABIN8068916

Human GCK ELISA Kit, Colorimetric assay for quantification of Human GCK.

Quick Overview for GCK ELISA Kit (ABIN8068916)

Target: GCK (Glucokinase (Hexokinase 4) (GCK))

Binding Specificity: nitrated, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details GCK ELISA Kit

Purpose: Measures nitrated Glucokinase (GCK) and total Glucokinase in human cell and tissue lysates, enabling precise assessment of Glucokinase nitration 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-Glucokinase Antibody
• Anti-Nitration Detection Antibody
• Anti-Glucokinase 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 GCK

Target: GCK (Glucokinase (Hexokinase 4) (GCK))

Alternative Name: Glucokinase

Background:

Gene Symbols: GCK

Background: The Nitrated-Glucokinase and Total-Glucokinase ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human nitrated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan Glucokinase antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Glucokinase present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-nitration antibody is used to detect nitrated Glucokinase or biotinylated anti-Glucokinase antibody is used to detect total Glucokinase . 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 nitrated Glucokinase or total Glucokinase 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): Hexokinase-4 (HK4), Glucokinase, Hexokinase type IV (HK IV), Hexokinase-D

Gene ID: 2645

UniProt: P35557

Pathways: MAPK Signaling, Positive Regulation of Peptide Hormone Secretion, Carbohydrate Homeostasis, Cellular Glucan Metabolic Process, Regulation of Carbohydrate Metabolic Process