Human Cell Biology ELISA Kits 1
Human LOX-1 (Lectin Like Oxidized Low Density Lipoprotein Receptor 1) ELISA Kit (HUES02637)
- SKU:
- HUES02637
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P78380
- Sensitivity:
- 0.09ng/mL
- Range:
- 0.16-10ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- Oxidized low-density lipoprotein receptor 1,Ox-LDL receptor 1, lectin-type oxidized LDL receptor 1,LOX-1,OLR1, CLEC8A, LOX1, LOXIN, SCARE1, SLOX1
- 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: | Colormetric |
Detection Range: | 0.16-10 ng/mL |
Sensitivity: | 0.10 ng/mL |
Sample Volume Required Per Well: | 100µL |
Sample Type: | Serum, plasma and other biological fluids |
Specificity: | This kit recognizes Human LOX-1 in samples. No significant cross-reactivity or interference between Human LOX-1 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 LOX-1. 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 LOX-1 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 LOX-1, 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 LOX-1. The concentration of Human LOX-1 in samples can be calculated by comparing the OD of the samples to the standard curve.
UniProt Protein Function: | OLR1: Receptor that mediates the recognition, internalization and degradation of oxidatively modified low density lipoprotein (oxLDL) by vascular endothelial cells. OxLDL is a marker of atherosclerosis that induces vascular endothelial cell activation and dysfunction, resulting in pro-inflammatory responses, pro- oxidative conditions and apoptosis. Its association with oxLDL induces the activation of NF-kappa-B through an increased production of intracellular reactive oxygen and a variety of pro- atherogenic cellular responses including a reduction of nitric oxide (NO) release, monocyte adhesion and apoptosis. In addition to binding oxLDL, it acts as a receptor for the HSP70 protein involved in antigen cross-presentation to naive T-cells in dendritic cells, thereby participating in cell-mediated antigen cross-presentation. Also involved in inflammatory process, by acting as a leukocyte-adhesion molecule at the vascular interface in endotoxin-induced inflammation. Also acts as a receptor for advanced glycation end (AGE) products, activated platelets, monocytes, apoptotic cells and both Gram-negative and Gram- positive bacteria. Independent association genetic studies have implicated OLR1 gene variants in myocardial infarction susceptibility. OLR1 may be involved in Alzheimer disease (AD). Involvement in AD is however unclear: according to some authors (PubMed:12354387, PubMed:12810610 and PubMed:15976314), variations in OLR1 modify the risk of AD, while according to other (PubMed:15000751 and PubMed:15060104) they do not. 2 isoforms of the human protein are produced by alternative splicing. |
UniProt Protein Details: | Protein type:Membrane protein, integral; Receptor, misc. Chromosomal Location of Human Ortholog: 12p13. 2-p12. 3 Cellular Component: nucleoplasm; membrane; intracellular membrane-bound organelle; integral to plasma membrane; plasma membrane; receptor complex; lipid raft Molecular Function:low-density lipoprotein receptor activity; protein binding; carbohydrate binding Biological Process: cell death; receptor-mediated endocytosis; leukocyte adhesion; response to hydrogen peroxide; blood circulation; lipoprotein metabolic process; inflammatory response; proteolysis; blood coagulation; leukocyte migration Disease: Myocardial Infarction, Susceptibility To |
NCBI Summary: | This gene encodes a low density lipoprotein receptor that belongs to the C-type lectin superfamily. This gene is regulated through the cyclic AMP signaling pathway. The encoded protein binds, internalizes and degrades oxidized low-density lipoprotein. This protein may be involved in the regulation of Fas-induced apoptosis. This protein may play a role as a scavenger receptor. Mutations of this gene have been associated with atherosclerosis, risk of myocardial infarction, and may modify the risk of Alzheimer's disease. Alternate splicing results in multiple transcript variants. [provided by RefSeq, Feb 2010] |
UniProt Code: | P78380 |
NCBI GenInfo Identifier: | 73621335 |
NCBI Gene ID: | 4973 |
NCBI Accession: | P78380. 1 |
UniProt Secondary Accession: | P78380,Q2PP00, Q7Z484, A8K7V9, B4DI48, G3V1I4, |
UniProt Related Accession: | P78380 |
Molecular Weight: | 21,425 Da |
NCBI Full Name: | Oxidized low-density lipoprotein receptor 1 |
NCBI Synonym Full Names: | oxidized low density lipoprotein (lectin-like) receptor 1 |
NCBI Official Symbol: | OLR1 |
NCBI Official Synonym Symbols: | LOX1; LOXIN; SLOX1; CLEC8A; SCARE1 |
NCBI Protein Information: | oxidized low-density lipoprotein receptor 1; hLOX-1; ox LDL receptor 1; lectin-type oxidized LDL receptor 1; scavenger receptor class E, member 1; C-type lectin domain family 8 member A; oxidized low-density lipoprotein receptor 1, soluble form |
UniProt Protein Name: | Oxidized low-density lipoprotein receptor 1 |
UniProt Synonym Protein Names: | C-type lectin domain family 8 member A; Lectin-like oxidized LDL receptor 1; LOX-1; Lectin-like oxLDL receptor 1; hLOX-1; Lectin-type oxidized LDL receptor 1Cleaved into the following chain:Oxidized low-density lipoprotein receptor 1, soluble form |
Protein Family: | Oxidized low-density lipoprotein receptor |
UniProt Gene Name: | OLR1 |
UniProt Entry Name: | OLR1_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 (ng/mL) | O.D | Average | Corrected |
10 | 2.391 2.429 | 2.41 | 2.346 |
5 | 1.579 1.635 | 1.607 | 1.543 |
2.5 | 0.959 0.937 | 0.948 | 0.884 |
1.25 | 0.53 0.532 | 0.531 | 0.467 |
0.63 | 0.295 0.279 | 0.287 | 0.223 |
0.32 | 0.187 0.159 | 0.173 | 0.109 |
0.16 | 0.112 0.128 | 0.12 | 0.056 |
0 | 0.057 0.071 | 0.064 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human LOX-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human LOX-1 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 (ng/mL) | 0.53 | 1.43 | 3.40 | 0.56 | 1.31 | 3.29 |
Standard deviation | 0.04 | 0.06 | 0.12 | 0.03 | 0.07 | 0.17 |
C V (%) | 7.55 | 4.20 | 3.53 | 5.36 | 5.34 | 5.17 |
Recovery
The recovery of Human LOX-1 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) | 93-107 | 100 |
EDTA plasma (n=5) | 89-101 | 96 |
Cell culture media (n=5) | 91-103 | 97 |
Linearity
Samples were spiked with high concentrations of Human LOX-1 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 (%) | 98-112 | 89-101 | 91-107 |
Average (%) | 104 | 94 | 98 | |
1:4 | Range (%) | 93-108 | 85-99 | 83-93 |
Average (%) | 99 | 90 | 88 | |
1:8 | Range (%) | 91-102 | 79-92 | 84-99 |
Average (%) | 96 | 86 | 91 | |
1:16 | Range (%) | 92-107 | 84-98 | 87-102 |
Average (%) | 100 | 91 | 93 |
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.