Mouse Cell Signalling ELISA Kits 2
Mouse LOX (Lysyl Oxidase) CLIA Kit (MOES00005)
- SKU:
- MOES00005
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 37.5pg/mL
- Range:
- 62.5-4000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- Protein-Lysine 6-Oxidase, MGC105112
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection method: | Chemiluminescence |
| Detection range: | 62.50-4000 pg/mL |
| Sensitivity: | 37.50 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Mouse LOX in samples. No significant cross-reactivity or interference between Mouse LOX 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 Mouse LOX. 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 Mouse LOX 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 Mouse LOX, 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 Mouse LOX. The concentration of Mouse LOX in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | LOX: Responsible for the post-translational oxidative deamination of peptidyl lysine residues in precursors to fibrous collagen and elastin. In addition to cross-linking of extracellular matrix proteins, may have a direct role in tumor suppression. Belongs to the lysyl oxidase family. |
| UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide; EC 1. 4. 3. 13; Oxidoreductase Cellular Component: proteinaceous extracellular matrix; collagen; extracellular region; nucleus Molecular Function:protein binding; copper ion binding; protein-lysine 6-oxidase activity; metal ion binding; oxidoreductase activity; oxidoreductase activity, acting on the CH-NH2 group of donors, oxygen as acceptor Biological Process: blood vessel development; elastic fiber assembly; collagen fibril organization; wound healing; response to steroid hormone stimulus; response to hormone stimulus; lung development |
| NCBI Summary: | This gene encodes a precursor protein that belongs to the lysyl oxidase family of proteins. The secreted proprotein is proteolytically processed to an active mature peptide and a propeptide. This propeptide is thought to function in tumor suppression by inhibiting the Ras signaling pathway. The active enzyme plays a role in cross-linking of collagen and elastin and is essential for development of cardiovascular and respiratory systems, and development of skin and connective tissue. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Oct 2013] |
| UniProt Code: | P28301 |
| NCBI GenInfo Identifier: | 555943929 |
| NCBI Gene ID: | 16948 |
| NCBI Accession: | NP_001273110. 1 |
| UniProt Related Accession: | P28301 |
| Molecular Weight: | Predicted isoelectric point: 6. 1Predicted Molecular Mass: 55. 4kDaAccurate Molecular Mass: 54kDa as determined by SDS-PAGE reducing conditions. |
| NCBI Full Name: | protein-lysine 6-oxidase preproprotein |
| NCBI Synonym Full Names: | lysyl oxidase |
| NCBI Official Symbol: | Lox |
| NCBI Official Synonym Symbols: | rrg; TSC-160; AI893619 |
| NCBI Protein Information: | protein-lysine 6-oxidase; ras recision; ras excision protein |
| UniProt Protein Name: | Protein-lysine 6-oxidase |
| UniProt Synonym Protein Names: | Lysyl oxidase; Ras excision protein |
| Protein Family: | Linoleate 9/13-lipoxygenase |
| UniProt Gene Name: | Lox |
| UniProt Entry Name: | LYOX_MOUSE |
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 |
| 4000 | 58177 59985 | 59081 | 59046 |
| 2000 | 27854 27864 | 27859 | 27824 |
| 1000 | 14589 12379 | 13484 | 13449 |
| 500 | 6518 6692 | 6605 | 6570 |
| 250 | 3419 3067 | 3243 | 3208 |
| 125 | 1599 1563 | 1581 | 1546 |
| 62.50 | 724 786 | 755 | 720 |
| 0 | 33 37 | 35 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse LOX were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse LOX 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) | 198.56 | 563.17 | 1475.33 | 207.04 | 570.40 | 1387.73 |
| Standard deviation | 19.36 | 64.37 | 97.08 | 19.25 | 57.67 | 89.23 |
| C V (%) | 9.75 | 11.43 | 6.58 | 9.30 | 10.11 | 6.43 |
Recovery
The recovery of Mouse LOX 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) | 85-98 | 91 |
| EDTA plasma (n=5) | 93-104 | 99 |
| Cell culture media (n=5) | 86-99 | 91 |
Linearity
Samples were spiked with high concentrations of Mouse LOX 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 (%) | 90-104 | 84-96 | 99-112 |
| Average (%) | 95 | 90 | 106 | |
| 1:4 | Range (%) | 85-98 | 97-114 | 94-111 |
| Average (%) | 92 | 105 | 102 | |
| 1:8 | Range (%) | 89-106 | 90-105 | 87-98 |
| Average (%) | 97 | 98 | 93 | |
| 1:16 | Range (%) | 99-116 | 83-95 | 89-104 |
| Average (%) | 106 | 90 | 95 |
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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