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AB125967

Human Glucokinase ELISA Kit (GCK)

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Human Glucokinase ELISA Kit (GCK) is a sandwich ELISA designed to quantify Human Glucokinase (GCK) with a sensitivity of 0.32 ng/mL.

- Colorimetric sandwich ELISA - 450 nm readout - works on any plate reader
- Wide dynamic range - quantifies 0.78 - 12.5 ng/mL

View Alternative Names

Hexokinase-4, HK4, Glucokinase, Hexokinase type IV, Hexokinase-D, HK IV, GCK

1 Images
Sandwich ELISA - Human Glucokinase ELISA Kit (GCK) (AB125967)
  • sELISA

Supplier Data

Sandwich ELISA - Human Glucokinase ELISA Kit (GCK) (AB125967)

Representative Standard Curve using ab125967

Key facts

Detection method

Colorimetric

Sample types

Plasma, Cell Lysate, Serum, Tissue

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 0.32 ng/mL

Range

0.391 - 25 ng/mL

Assay time

4h

Assay Platform

Microplate

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Primary Antibodies

AB88056

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human Glucokinase ELISA Kit (GCK) ab125967 is a sandwich ELISA to measure Human Glucokinase (GCK) in serum, plasma, cell lysates and tissue samples with a sensitivity of 0.32 ng/ml.

How the assay works

A Glucokinase specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently a Glucokinase specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Conjugate is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize Streptavidin-Peroxidase enzymatic reaction. TMB is catalyzed by Streptavidin-Peroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the amount of Glucokinase captured in plate.
The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.

Assay Specificity

Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility. Please check the protocol booklet for more details

Human Glucokinase ELISA Kit (GCK) ab125967 protocol summary

1. Add standard or sample to appropriate wells. Incubate at room temperature
2. Wash and add prepared biotin antibody to each well. Incubate at room temperature.
3. Wash and add prepared Streptavidin-Peroxidase Conjugate. Incubate at room temperature
4. Add Chromogen Substrate to each well. Incubate at room temperature
5. Add Stop Solution to each well. Read at 450 nm immediately.
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Precision

[ { "reproducibilityType": "Intra", "sample": "Overall", "replicates": 0, "mean": null, "standardDeviation": null, "coefficientOfVariability": "5.4" }, { "reproducibilityType": "Inter", "sample": "Overall", "replicates": 0, "mean": null, "standardDeviation": null, "coefficientOfVariability": "10.2" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
Multi
Storage information
Please refer to protocols
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Glucokinase also known as GCK is an enzyme with a mass around 50 kDa. It functions by catalyzing the phosphorylation of glucose to glucose-6-phosphate an important step in glycolysis. Glucokinase displays a higher affinity for glucose as compared to other hexokinases which is indicated by its specific kinetic property known as the Km value. This enzyme is mainly expressed in the liver and pancreatic beta-cells where it plays an important role in glucose metabolism. Additionally glucokinase can also be found in the hypothalamus indicating its broader reach in carbohydrate sensing.
Biological function summary

Glucokinase acts as a glucose sensor regulating blood sugar levels and insulin secretion. This process does not involve the enzyme as part of a larger complex. Instead glucokinase functions independently to sense glucose concentrations and adjusts the rate of glucose phosphorylation accordingly. Its activity significantly affects the balance of glucose uptake and energy production especially during postprandial states when glucose levels rise. Glucokinase activity is therefore important to maintaining glucose homeostasis.

Pathways

Glucokinase plays a central role in the glycolytic pathway and interacts with the insulin signaling pathway. Within these pathways it influences how the liver stores glucose as glycogen and controls insulin release from pancreatic beta-cells. Glucokinase activity is modulated by glucose levels and is closely related to insulin receptor substrates that affect downstream insulin signaling. Its function is important for the proper regulation of these pathways linking glucose sensing with metabolic control in varied tissues.

Glucokinase is pivotal in the context of diabetes mellitus and hyperglycemia. Mutations or dysregulation of glucokinase can lead to different forms of diabetes including maturity-onset diabetes of the young type 2 (MODY2). Glucokinase inhibitors and activators are being explored as potential therapeutic agents for modulating glucose levels in diabetics. Additionally its role interfaces with the functions of insulin tying glucokinase closely to conditions of insulin resistance and pancreatic dysfunction. Understanding these connections is instrumental for developing targeted treatment strategies.
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Product protocols

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Target data

Catalyzes the phosphorylation of hexose, such as D-glucose, D-fructose and D-mannose, to hexose 6-phosphate (D-glucose 6-phosphate, D-fructose 6-phosphate and D-mannose 6-phosphate, respectively) (PubMed : 11916951, PubMed : 15277402, PubMed : 17082186, PubMed : 18322640, PubMed : 19146401, PubMed : 25015100, PubMed : 7742312, PubMed : 8325892). Compared to other hexokinases, has a weak affinity for D-glucose, and is effective only when glucose is abundant (By similarity). Mainly expressed in pancreatic beta cells and the liver and constitutes a rate-limiting step in glucose metabolism in these tissues (PubMed : 11916951, PubMed : 15277402, PubMed : 18322640, PubMed : 25015100, PubMed : 8325892). Since insulin secretion parallels glucose metabolism and the low glucose affinity of GCK ensures that it can change its enzymatic activity within the physiological range of glucose concentrations, GCK acts as a glucose sensor in the pancreatic beta cell (By similarity). In pancreas, plays an important role in modulating insulin secretion (By similarity). In liver, helps to facilitate the uptake and conversion of glucose by acting as an insulin-sensitive determinant of hepatic glucose usage (By similarity). Required to provide D-glucose 6-phosphate for the synthesis of glycogen (PubMed : 8878425). Mediates the initial step of glycolysis by catalyzing phosphorylation of D-glucose to D-glucose 6-phosphate (PubMed : 7742312).
See full target information Hexokinase-4
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websiteProtocolBooklet
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Product promise

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