Mouse LRP-1 (Low-Density Lipoprotein-Receptor-Related Protein) ELISA Kit (MOES01745)
Mouse LRP-1 (Low-Density Lipoprotein-Receptor-Related Protein) ELISA Kit
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 Mouse LRP-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 Mouse LRP-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 Mouse LRP-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 Mouse LRP-1. The concentration of Mouse LRP-1 in samples can be calculated by comparing the OD of the samples to the standard curve.
|Detection Range||0.63-40 ng/mL|
|Sample Volume Required Per Well||100uL|
|Sample Type||Serum, plasma and other biological fluids|
This kit recognizes Mouse LRP-1 in samples. No significant cross-reactivity or interference between Mouse LRP-1 and analogues was observed.
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse LRP-1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse LRP-1 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.86||5.31||3.21||5.83||4.54||4.65|
The recovery of Mouse LRP-1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||90-102||96|
|Cell culture media (n=5)||93-107||99|
Samples were spiked with high concentrations of Mouse LRP-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)|
Kit Components & Storage
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.
|Micro ELISA Plate(Dismountable)||8 wells X12 strips||-20'C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100X)||1 vial, 120 uL|
|Concentrated HRP Conjugate (100X)||1 vial, 120 uL||-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 (25X)||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|
Mouse LRP-1 (Low-Density Lipoprotein-Receptor-Related Protein) ELISA Kit (MOES01745) Assay procedure
- 1. 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.
- 2. Aliquot 100µl of standard solutions into the standard wells.
- 3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- 4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- 5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- 6. 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.
- 7. 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.
- 8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- 9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 10. 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.
- 11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- 12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
Mouse LRP-1 (Low-Density Lipoprotein-Receptor-Related Protein) ELISA Kit (MOES01745) Protein Information
|UniProt Protein Function:||LRP1: a receptor involved in endocytosis, phagocytosis of apoptotic cells, and neural signaling. Modulates cellular events including APP metabolism, kinase-dependent intracellular signaling, neuronal calcium signaling and neurotransmission. Involved in the plasma clearance of chylomicron remnants and activated alpha 2-macroglobulin, as well as the local metabolism of complexes between plasminogen activators and their endogenous inhibitors. Plays a fundamental role in cellular signaling pathways in the brain. Mediates neural NMDA gating via a complex of LRP1, PSD-95 and NMDAR. A type I membrane protein. After cleavage, the intracellular domain (LRPICD) is detected both in the cytoplasm and in the nucleus. Most abundant in liver, brain and lung.|
|UniProt Protein Details:|
Protein type:Cell surface; Membrane protein, integral; Motility/polarity/chemotaxis; Receptor, misc.
Chromosomal Location of Human Ortholog: 10|10 D3
Cellular Component: basolateral plasma membrane; cell soma; clathrin-coated pit; clathrin-coated vesicle; cytoplasm; cytosol; dendrite; early endosome; endosome; focal adhesion; integral component of membrane; integral component of plasma membrane; lysosomal membrane; membrane; nucleolus; nucleus; perinuclear region of cytoplasm; plasma membrane; receptor complex
Molecular Function:alpha-2 macroglobulin receptor activity; apolipoprotein binding; calcium ion binding; clathrin heavy chain binding; metal ion binding; protease binding; protein binding; protein complex binding; RNA binding
Biological Process: apoptotic cell clearance; astrocyte activation during immune response; cardiac septum development; cell proliferation; cellular response to amyloid-beta; cholesterol metabolic process; endocytosis; multicellular organism development; negative regulation of neuron apoptosis; negative regulation of neuron projection development; negative regulation of platelet-derived growth factor receptor-beta signaling pathway; negative regulation of smooth muscle cell migration; negative regulation of Wnt signaling pathway; positive regulation of amyloid-beta clearance; positive regulation of cell death; positive regulation of cholesterol efflux; positive regulation of endocytosis; positive regulation of lipid transport; positive regulation of lysosomal protein catabolic process; positive regulation of protein binding; positive regulation of protein catabolic process; positive regulation of protein transport; protein kinase C-activating G-protein coupled receptor signaling pathway; receptor-mediated endocytosis; regulation of actin cytoskeleton organization; regulation of cholesterol transport; regulation of phospholipase A2 activity; regulation of protein metabolic process; transcytosis
|NCBI GenInfo Identifier:||124494256|
|NCBI Gene ID:||16971|
|UniProt Secondary Accession:||Q91ZX7,Q61291, Q920Y4,|
|UniProt Related Accession:||Q91ZX7|
|Molecular Weight:||504,742 Da|
|NCBI Full Name:||prolow-density lipoprotein receptor-related protein 1|
|NCBI Synonym Full Names:||low density lipoprotein receptor-related protein 1|
|NCBI Official Symbol:||Lrp1|
|NCBI Official Synonym Symbols:||Lrp; A2mr; CD91; AI316852; b2b1554Clo|
|NCBI Protein Information:||prolow-density lipoprotein receptor-related protein 1|
|UniProt Protein Name:||Prolow-density lipoprotein receptor-related protein 1|
|UniProt Synonym Protein Names:||Alpha-2-macroglobulin receptor; A2MR|
|Protein Family:||Low-density lipoprotein receptor-related protein|
|UniProt Gene Name:||Lrp1|