Description
Human GP39 (Glycoprotein 39, Cartilage) ELISA Kit
The Human GP39 (Glycoprotein-39) ELISA Kit is a highly sensitive and specific assay designed for the accurate detection of GP39 levels in human serum, plasma, and cell culture supernatants. This kit offers reliable and reproducible results, making it ideal for a variety of research applications.GP39, also known as Chitinase 3-like 1 (CHI3L1), is a glycoprotein that plays a key role in inflammation, tissue remodeling, and extracellular matrix regulation.
It has been implicated in various diseases, including asthma, rheumatoid arthritis, and cancer, making it a valuable biomarker for studying these conditions and developing targeted therapies.With this ELISA kit, researchers can quantify GP39 levels with precision, enabling in-depth studies of its role in disease pathogenesis and potential therapeutic interventions. Invest in the Human GP39 ELISA Kit for accurate and insightful research findings.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 62.50-4000 pg/mL |
| Sensitivity: | 37.50 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human GP39 in samples. No significant cross-reactivity or interference between Human GP39 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 GP39. 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 GP39 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 GP39, 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 GP39. The concentration of Human GP39 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | CHI3L1: Carbohydrate-binding lectin with a preference for chitin. May play a role in defense against pathogens, or in tissue remodeling. May play an important role in the capacity of cells to respond to and cope with changes in their environment. A genetic variation in CHI3L1 is associated with susceptibility to asthma-related traits type 7 (ASRT7). Asthma-related traits include clinical symptoms of asthma, such as coughing, wheezing and dyspnea, bronchial hyperresponsiveness (BHR) as assessed by methacholine challenge test, serum IgE levels, atopy, and atopic dermatitis. Belongs to the glycosyl hydrolase 18 family. |
| UniProt Protein Details: | Protein type:Secreted, signal peptide; Secreted; Cell adhesion Chromosomal Location of Human Ortholog: 1q32. 1 Cellular Component: extracellular space; proteinaceous extracellular matrix; perinuclear region of cytoplasm; endoplasmic reticulum; cytoplasm Molecular Function:extracellular matrix structural constituent; chitin binding; chitinase activity; carbohydrate binding Biological Process: positive regulation of protein kinase B signaling cascade; positive regulation of angiogenesis; apoptosis; response to mechanical stimulus; cartilage development; carbohydrate metabolic process; activation of NF-kappaB-inducing kinase; immune response; chitin catabolic process; inflammatory response; lung development Disease: Schizophrenia; Asthma-related Traits, Susceptibility To, 7 |
| NCBI Summary: | Chitinases catalyze the hydrolysis of chitin, which is an abundant glycopolymer found in insect exoskeletons and fungal cell walls. The glycoside hydrolase 18 family of chitinases includes eight human family members. This gene encodes a glycoprotein member of the glycosyl hydrolase 18 family. The protein lacks chitinase activity and is secreted by activated macrophages, chondrocytes, neutrophils and synovial cells. The protein is thought to play a role in the process of inflammation and tissue remodeling. [provided by RefSeq, Sep 2009] |
| UniProt Code: | P36222 |
| NCBI GenInfo Identifier: | 84028186 |
| NCBI Gene ID: | 1116 |
| NCBI Accession: | P36222. 2 |
| UniProt Secondary Accession: | P36222,P30923, Q8IVA4, Q96HI7, B2R7B0, |
| UniProt Related Accession: | P36222 |
| Molecular Weight: | 42,625 Da |
| NCBI Full Name: | Chitinase-3-like protein 1 |
| NCBI Synonym Full Names: | chitinase 3-like 1 (cartilage glycoprotein-39) |
| NCBI Official Symbol: | CHI3L1 |
| NCBI Official Synonym Symbols: | GP39; ASRT7; GP-39; YKL40; CGP-39; YKL-40; YYL-40; HC-gp39; HCGP-3P; hCGP-39 |
| NCBI Protein Information: | chitinase-3-like protein 1; 39 kDa synovial protein; cartilage glycoprotein 39 |
| UniProt Protein Name: | Chitinase-3-like protein 1 |
| UniProt Synonym Protein Names: | 39 kDa synovial protein; Cartilage glycoprotein 39; CGP-39; GP-39; hCGP-39; YKL-40 |
| Protein Family: | Chitinase-3-like protein |
| UniProt Gene Name: | CHI3L1 |
| UniProt Entry Name: | CH3L1_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 |
| 4000 | 2.328 2.358 | 2.343 | 2.284 |
| 2000 | 1.493 1.503 | 1.498 | 1.439 |
| 1000 | 0.845 0.829 | 0.837 | 0.778 |
| 500 | 0.42 0.428 | 0.424 | 0.365 |
| 250 | 0.226 0.198 | 0.212 | 0.153 |
| 125 | 0.156 0.154 | 0.155 | 0.096 |
| 62.50 | 0.106 0.112 | 0.109 | 0.05 |
| 0 | 0.051 0.067 | 0.059 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GP39 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human GP39 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) | 195.87 | 424.36 | 1816.59 | 190.47 | 437.11 | 1884.97 |
| Standard deviation | 12.85 | 18.16 | 84.65 | 12.38 | 23.25 | 67.86 |
| C V (%) | 6.56 | 4.28 | 4.66 | 6.50 | 5.32 | 3.60 |
Recovery
The recovery of Human GP39 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) | 93-105 | 99 |
| EDTA plasma (n=5) | 96-106 | 101 |
| Cell culture media (n=5) | 89-105 | 96 |
Linearity
Samples were spiked with high concentrations of Human GP39 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 (%) | 86-102 | 87-101 | 96-110 |
| Average (%) | 93 | 93 | 103 | |
| 1:4 | Range (%) | 91-101 | 86-98 | 88-99 |
| Average (%) | 96 | 91 | 94 | |
| 1:8 | Range (%) | 87-100 | 86-102 | 85-98 |
| Average (%) | 92 | 93 | 90 | |
| 1:16 | Range (%) | 86-98 | 84-95 | 83-97 |
| Average (%) | 93 | 90 | 89 |
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 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL 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 100 µL 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 350 µL 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 100 µL 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 90 µL 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.
