null

Mouse GCK / Glucokinase ELISA Kit

SKU:
MOFI00271
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P52792
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
Gck, Glucokinase, FGQTL3, GLK, Hexokinase-4, Hexokinase-D, HHF3, HK4, HXKP, LGLK, GK, glucokinase, hexokinase 4, HKIV, maturity onset diabetes of the young 2, MODY2
Reactivity:
Mouse
Research Area:
Metabolism
€649
Frequently bought together:

Description

system_update_altDatasheetsystem_update_altMSDS

Mouse GCK/ Glucokinase ELISA Kit

The Mouse GCK (Glucokinase) ELISA Kit is a powerful tool for the precise quantification of glucokinase levels in mouse serum, plasma, and cell culture supernatants. With excellent sensitivity and specificity, this kit ensures accurate and consistent results, making it ideal for various research purposes.Glucokinase is a key enzyme that plays a vital role in glucose metabolism, regulating glucose uptake and utilization in the body. Dysregulation of glucokinase has been associated with metabolic disorders such as diabetes and obesity, highlighting its importance as a potential therapeutic target.

By accurately measuring glucokinase levels, researchers can gain valuable insights into metabolic pathways and potential therapeutic interventions for metabolic disorders. The Mouse GCK ELISA Kit provides a reliable and efficient tool for studying glucokinase and its role in metabolic health.

Product Name:Mouse GCK / Glucokinase ELISA Kit
Product Code:MOFI00271
Size:96 Assays
Alias:Gck, Glucokinase, FGQTL3, GLK, Hexokinase-4, Hexokinase-D, HHF3, HK4, HXKP, LGLK, GK, glucokinase, hexokinase 4, HKIV, maturity onset diabetes of the young 2, MODY2
Detection Method:Sandwich ELISA
Application:This immunoassay kit allows for the in vitro quantitative determination of Mouse Gck concentrations in serum plasma and other biological fluids.
Sensitivity:0.094ng/ml
Range:0.156-10ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery: Matrices listed below were spiked with certain level of Mouse Gck and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse Gck in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)87-9994
EDTA plasma(n=5)92-10599
UFH plasma(n=5)86-10393
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse Gck and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)91-103%85-104%92-105%
EDTA plasma(n=5)92-100%84-98%83-99%
UFH plasma(n=5)80-94%81-90%81-100%
Intra Assay:CV <8%
Inter Assay:CV <10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8-12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP52792
UniProt Protein Function:GK: a glycolytic enzyme that catalyzes glucose metabolism in the liver and pancreatic beta cells. Acts as a ""glucose sensor"" in beta cells. The first and rate-limiting step in glycosis, a pathway that produces energy in the form of ATP from glucose. Glucokinase traps glucose inside the cell by catalyzing its phosphorylation to produce glucose-6-phosphate. Has a lower affinity for glucose than the three other isozymes of hexokinase, allowing other organs such as the brain and muscles to have first call on glucose when its supply is limited. Unlike other hexokinases, glucokinase is not inhibited by glucose-6-phosphate. Glucokinase is found in the outer membrane compartment of mitochondria. May bind VDAC, suppressing mitochondrial function. Glucokinase transcription is induced by insulin, perhaps via the activation of Stat 5B. Mutant glucokinase causes a rare form of diabetes and may also play a role in type 2 diabetes. Three splice variant isoforms of human glucokinase have been described.
UniProt Protein Details:

Protein type:Carbohydrate Metabolism - amino sugar and nucleotide sugar; Kinase, other; Carbohydrate Metabolism - galactose; Mitochondrial; Carbohydrate Metabolism - glycolysis and gluconeogenesis; EC 2.7.1.2; Carbohydrate Metabolism - starch and sucrose

Cellular Component: nucleoplasm; mitochondrion; cell; cytoplasm; actin filament; cell cortex; cytosol; basal cortex; nucleus; secretory granule

Molecular Function:transferase activity; hexokinase activity; protein binding; magnesium ion binding; nucleotide binding; kinase activity; ADP binding; phosphotransferase activity, alcohol group as acceptor; glucokinase activity; protein phosphatase binding; ATP binding; glucose binding

Biological Process: positive regulation of glycogen biosynthetic process; glucose 6-phosphate metabolic process; lipid homeostasis; glucose homeostasis; NADP metabolic process; cellular response to glucose starvation; elevation of cytosolic calcium ion concentration; detection of glucose; negative regulation of gluconeogenesis; second-messenger-mediated signaling; glycogen biosynthetic process; glycolysis; regulation of potassium ion transport; glucose metabolic process; positive regulation of insulin secretion; fructose 2,6-bisphosphate metabolic process; cellular response to insulin stimulus; carbohydrate metabolic process; carbohydrate phosphorylation; positive regulation of glycolysis; negative regulation of epinephrine secretion; cell glucose homeostasis; regulation of insulin secretion; phosphorylation; positive regulation of phosphorylation

UniProt Code:P52792
NCBI GenInfo Identifier:31982798
NCBI Gene ID:103988
NCBI Accession:NP_034422.2
UniProt Secondary Accession:P52792,P52791,
UniProt Related Accession:P52792
Molecular Weight:52,089 Da
NCBI Full Name:glucokinase
NCBI Synonym Full Names:glucokinase
NCBI Official Symbol:Gck  
NCBI Official Synonym Symbols:Gk; GLK; HK4; HKIV; HXKP; MODY2; Gls006  
NCBI Protein Information:glucokinase; HK IV; hexokinase 4; hexokinase-4; hexokinase-D; hexokinase type IV
UniProt Protein Name:Glucokinase
UniProt Synonym Protein Names:Hexokinase type IV; HK IV; Hexokinase-4; HK4; Hexokinase-D
Protein Family:Glucokinase
UniProt Gene Name:Gck  
UniProt Entry Name:HXK4_MOUSE

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

StepProcedure
1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample (Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12. Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum:

If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

Plasma:

Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 - g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.

Urine & Cerebrospinal Fluid:

Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.

Cell culture supernatant:

Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.

Cell lysates:

Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20°C.

Tissue homogenates:

The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.

Tissue lysates:

Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.

Breast Milk:

Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

Related Products