Human Cyr61 (Cysteine Rich Protein, Angiogenic Inducer 61) CLIA Kit (HUES00738)
- SKU:
- HUES00738
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 9.38pg/mL
- Range:
- 15.63-1000pg/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human Cyr61 in samples. No significant cross-reactivity or interference between Human Cyr61 and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human Cyr61. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human Cyr61 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 Cyr61, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human Cyr61. The concentration of Human Cyr61 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | CYR61: Promotes cell proliferation, chemotaxis, angiogenesis and cell adhesion. Appears to play a role in wound healing by up- regulating, in skin fibroblasts, the expression of a number of genes involved in angiogenesis, inflammation and matrix remodeling including VEGA-A, VEGA-C, MMP1, MMP3, TIMP1, uPA, PAI-1 and integrins alpha-3 and alpha-5. CYR61-mediated gene regulation is dependent on heparin-binding. Down-regulates the expression of alpha-1 and alpha-2 subunits of collagen type-1. Promotes cell adhesion and adhesive signaling through integrin alpha-6/beta-1, cell migration through integrin alpha-v/beta-5 and cell proliferation through integrin alpha-v/beta-3. Belongs to the CCN family. |
| UniProt Protein Details: | Protein type:Secreted, signal peptide; Secreted Chromosomal Location of Human Ortholog: 1p22. 3 Cellular Component: extracellular matrix; proteinaceous extracellular matrix Molecular Function:heparin binding; integrin binding; insulin-like growth factor binding; extracellular matrix binding Biological Process: anatomical structure morphogenesis; extracellular matrix organization and biogenesis; positive regulation of caspase activity; chemotaxis; intussusceptive angiogenesis; osteoblast differentiation; cell proliferation; positive regulation of osteoblast differentiation; cell-cell signaling; positive regulation of protein kinase activity; positive regulation of osteoblast proliferation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of protein amino acid phosphorylation; regulation of cell growth; cell adhesion; positive regulation of BMP signaling pathway; positive regulation of cell migration; negative regulation of apoptosis |
| NCBI Summary: | The secreted protein encoded by this gene is growth factor-inducible and promotes the adhesion of endothelial cells. The encoded protein interacts with several integrins and with heparan sulfate proteoglycan. This protein also plays a role in cell proliferation, differentiation, angiogenesis, apoptosis, and extracellular matrix formation. [provided by RefSeq, Sep 2011] |
| UniProt Code: | O00622 |
| NCBI GenInfo Identifier: | 3023601 |
| NCBI Gene ID: | 3491 |
| NCBI Accession: | O00622. 1 |
| UniProt Secondary Accession: | O00622,O14934, O43775, Q9BZL7, |
| UniProt Related Accession: | O00622 |
| Molecular Weight: | |
| NCBI Full Name: | Protein CYR61 |
| NCBI Synonym Full Names: | cysteine-rich, angiogenic inducer, 61 |
| NCBI Official Symbol: | CYR61 |
| NCBI Official Synonym Symbols: | CCN1; GIG1; IGFBP10 |
| NCBI Protein Information: | protein CYR61; IBP-10; IGFBP-10; CCN family member 1; IGF-binding protein 10; cysteine-rich, anigogenic inducer, 61; cysteine-rich heparin-binding protein 61; insulin-like growth factor-binding protein 10 |
| UniProt Protein Name: | Protein CYR61 |
| UniProt Synonym Protein Names: | CCN family member 1; Cysteine-rich angiogenic inducer 61; Insulin-like growth factor-binding protein 10; IBP-10; IGF-binding protein 10; IGFBP-10; Protein GIG1 |
| Protein Family: | Protein |
| UniProt Gene Name: | CYR61 |
| UniProt Entry Name: | CYR61_HUMAN |
As the RLU 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) | RLU | Average | Corrected |
| 1000 | 52876 54474 | 53675 | 53650 |
| 500 | 22256 22722 | 22489 | 22464 |
| 250 | 10318 10104 | 10211 | 10186 |
| 125 | 4679 5123 | 4901 | 4876 |
| 62.5 | 2574 2332 | 2453 | 2428 |
| 31.25 | 1376 1186 | 1281 | 1256 |
| 15.63 | 673 743 | 708 | 683 |
| 0 | 25 25 | 25 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human Cyr61 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human Cyr61 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) | 45.48 | 95.44 | 402.47 | 48.18 | 93.11 | 393.99 |
| Standard deviation | 4.97 | 7.42 | 24.27 | 4.48 | 7.09 | 30.97 |
| C V (%) | 10.93 | 7.77 | 6.03 | 9.30 | 7.61 | 7.86 |
Recovery
The recovery of Human Cyr61 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) | 90-105 | 97 |
| EDTA plasma (n=5) | 91-107 | 99 |
| Cell culture media (n=5) | 92-109 | 100 |
Linearity
Samples were spiked with high concentrations of Human Cyr61 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 (%) | 102-119 | 92-107 | 95-108 |
| Average (%) | 109 | 98 | 103 | |
| 1:4 | Range (%) | 97-111 | 88-98 | 89-101 |
| Average (%) | 105 | 93 | 95 | |
| 1:8 | Range (%) | 100-118 | 89-101 | 97-108 |
| Average (%) | 108 | 94 | 103 | |
| 1:16 | Range (%) | 100-114 | 85-99 | 98-112 |
| Average (%) | 106 | 92 | 105 |
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 CLIA 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 A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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