Description
Human GPI (Glucose 6 Phosphate Isomerase) ELISA Kit
The Human GPI (Glucose-6-Phosphate Isomerase) ELISA Kit is specifically designed for the accurate quantification of GPI levels in human serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, ensuring robust and reproducible results for a variety of research applications.GPI is an important enzyme involved in glycolysis and gluconeogenesis pathways, playing a key role in glucose metabolism. Dysregulation of GPI has been implicated in various diseases such as cancer, diabetes, and autoimmune disorders, highlighting its potential as a valuable biomarker for disease research and therapeutic development.
With the Human GPI ELISA Kit, researchers can confidently measure GPI levels in biological samples with precision and confidence, enabling deeper insights into the role of GPI in disease pathology and potential therapeutic interventions. Order now to advance your research with this cutting-edge ELISA kit.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 31.25-2000 pg/mL |
| Sensitivity: | 18.75 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human GPI in samples. No significant cross-reactivity or interference between Human GPI and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human GPI. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human GPI and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human GPI, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human GPI. The concentration of Human GPI in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | G6PI: belongs to the GPI family whose members encode multifunctional phosphoglucose isomerase proteins involved in energy pathways. A dimeric enzyme that catalyzes the reversible isomerization of glucose-6-phosphate and fructose-6-phosphate. Functions in different capacities inside and outside the cell. In the cytoplasm, the gene product is involved in glycolysis and gluconeogenesis, while outside the cell it functions as a neurotrophic factor for spinal and sensory neurons. Defects in this gene are the cause of nonspherocytic hemolytic anemia and a severe enzyme deficiency can be associated with hydrops fetalis, immediate neonatal death and neurological impairment. |
| UniProt Protein Details: | Protein type:Isomerase; Carbohydrate Metabolism - starch and sucrose; Carbohydrate Metabolism - amino sugar and nucleotide sugar; EC 5. 3. 1. 9; Carbohydrate Metabolism - glycolysis and gluconeogenesis; Cytokine; Carbohydrate Metabolism - pentose phosphate pathway; Apoptosis Chromosomal Location of Human Ortholog: 19q13. 1 Cellular Component: nucleoplasm; extracellular space; neuron projection; membrane; cytoplasm; plasma membrane; cytosol Molecular Function:monosaccharide binding; glucose-6-phosphate isomerase activity; growth factor activity; cytokine activity; intramolecular transferase activity Biological Process: aldehyde catabolic process; methylglyoxal biosynthetic process; glycolysis; glucose 6-phosphate metabolic process; glucose metabolic process; pathogenesis; negative regulation of caspase activity; humoral immune response; gluconeogenesis; learning and/or memory; hemostasis; carbohydrate metabolic process; negative regulation of neuron apoptosis; angiogenesis Disease: Hemolytic Anemia, Nonspherocytic, Due To Glucose Phosphate Isomerase Deficiency |
| NCBI Summary: | This gene encodes a member of the glucose phosphate isomerase protein family. The encoded protein has been identified as a moonlighting protein based on its ability to perform mechanistically distinct functions. In the cytoplasm, the gene product functions as a glycolytic enzyme (glucose-6-phosphate isomerase) that interconverts glucose-6-phophsate and fructose-6-phosphate. Extracellularly, the encoded protein (also referred to as neuroleukin) functions as a neurotrophic factor that promotes survival of skeletal motor neurons and sensory neurons, and as a lymphokine that induces immunoglobulin secretion. The encoded protein is also referred to as autocrine motility factor based on an additional function as a tumor-secreted cytokine and angiogenic factor. Defects in this gene are the cause of nonspherocytic hemolytic anemia and a severe enzyme deficiency can be associated with hydrops fetalis, immediate neonatal death and neurological impairment. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Jan 2014] |
| UniProt Code: | P06744 |
| NCBI GenInfo Identifier: | 17380385 |
| NCBI Gene ID: | 2821 |
| NCBI Accession: | P06744. 4 |
| UniProt Secondary Accession: | P06744,Q9BRD3, Q9BSK5, Q9UHE6, B4DG39, |
| UniProt Related Accession: | P06744 |
| Molecular Weight: | 558 |
| NCBI Full Name: | Glucose-6-phosphate isomerase |
| NCBI Synonym Full Names: | glucose-6-phosphate isomerase |
| NCBI Official Symbol: | GPI |
| NCBI Official Synonym Symbols: | AMF; NLK; PGI; PHI; GNPI; SA36; SA-36 |
| NCBI Protein Information: | glucose-6-phosphate isomerase; neuroleukin; oxoisomerase; sperm antigen 36; sperm antigen-36; phosphohexomutase; phosphosaccharomutase; phosphohexose isomerase; phosphoglucose isomerase; autocrine motility factor; hexosephosphate isomerase; glucose phosph |
| UniProt Protein Name: | Glucose-6-phosphate isomerase |
| UniProt Synonym Protein Names: | Autocrine motility factor; AMF; Neuroleukin; NLK; Phosphoglucose isomerase; PGI; Phosphohexose isomerase; PHI; Sperm antigen 36; SA-36 |
| Protein Family: | Glucose-6-phosphate isomerase |
| UniProt Gene Name: | GPI |
| UniProt Entry Name: | G6PI_HUMAN |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (pg/mL) | O.D | Average | Corrected |
| 2000 | 2.499 2.531 | 2.515 | 2.434 |
| 1000 | 1.62 1.624 | 1.622 | 1.541 |
| 500 | 0.952 0.918 | 0.935 | 0.854 |
| 250 | 0.514 0.538 | 0.526 | 0.445 |
| 125 | 0.297 0.273 | 0.285 | 0.204 |
| 62.5 | 0.198 0.17 | 0.184 | 0.103 |
| 31.25 | 0.126 0.142 | 0.134 | 0.053 |
| 0 | 0.078 0.084 | 0.081 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GPI were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human GPI were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 96.20 | 267.90 | 857.66 | 90.97 | 288.07 | 880.14 |
| Standard deviation | 6.02 | 13.21 | 39.28 | 4.74 | 16.62 | 42.86 |
| C V (%) | 6.26 | 4.93 | 4.58 | 5.21 | 5.77 | 4.87 |
Recovery
The recovery of Human GPI spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 90-103 | 97 |
| EDTA plasma (n=5) | 93-109 | 100 |
| Cell culture media (n=5) | 95-106 | 101 |
Linearity
Samples were spiked with high concentrations of Human GPI and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 85-99 | 89-100 | 98-112 |
| Average (%) | 92 | 95 | 105 | |
| 1:4 | Range (%) | 92-108 | 79-93 | 83-94 |
| Average (%) | 99 | 85 | 88 | |
| 1:8 | Range (%) | 90-100 | 83-94 | 87-102 |
| Average (%) | 95 | 88 | 93 | |
| 1:16 | Range (%) | 86-102 | 84-96 | 82-94 |
| Average (%) | 93 | 89 | 88 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
