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Human Prolow-density lipoprotein receptor-related protein 1 (LRP1) ELISA Kit

SKU:
HUEB2150
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q07954
Range:
0.156-10 ng/mL
ELISA Type:
Sandwich
Synonyms:
LRP1, Prolow-density lipoprotein receptor-related protein 1, LRP-1, Alpha-2-macroglobulin receptor, A2MR, Apolipoprotein E receptor, APOER, CD91
Reactivity:
Human
€649
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Description

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Product Name:Human Prolow-density lipoprotein receptor-related protein 1 (LRP1) ELISA Kit
Product Code:HUEB2150
Alias:Prolow-density lipoprotein receptor-related protein 1, LRP-1, Alpha-2-macroglobulin receptor, A2MR, Apolipoprotein E receptor, APOER, LRP1, A2MR, APR, CD91
Uniprot:Q07954
Reactivity:Human
Range:0.156-10 ng/mL
Detection Method:Sandwich
Size:96 Assay
Storage:Please see kit components below for exact storage details
Note:For research use only
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:Motility/polarity/chemotaxis; Cell surface; Membrane protein, integral; Receptor, misc.

Chromosomal Location of Human Ortholog: 12q13.3

Cellular Component: focal adhesion; cell soma; clathrin-coated vesicle; lysosomal membrane; integral to plasma membrane; dendrite; cytoplasm; plasma membrane; nucleolus; coated pit; endosome; receptor complex

Molecular Function:protein binding; lipoprotein transporter activity; protease binding; protein complex binding; apolipoprotein binding; calcium ion binding; receptor activity

Biological Process: positive regulation of protein transport; receptor-mediated endocytosis; cell proliferation; positive regulation of lipid transport; cholesterol metabolic process; phototransduction, visible light; regulation of phospholipase A2 activity; negative regulation of Wnt receptor signaling pathway; lipoprotein transport; regulation of actin cytoskeleton organization and biogenesis; protein kinase C activation; lipoprotein metabolic process; negative regulation of neuron apoptosis; negative regulation of smooth muscle cell migration; retinoid metabolic process; regulation of cholesterol transport; apoptotic cell clearance; aging

NCBI Summary:The protein encoded by this gene is an endocytic receptor involved in several cellular processes, including intracellular signaling, lipid homeostasis, and clearance of apoptotic cells. In addition, the encoded protein is necessary for the A2M-mediated clearance of secreted amyloid precursor protein and beta-amyloid, the main component of amyloid plaques found in Alzheimer patients. Expression of this gene decreases with age and has been found to be lower than controls in brain tissue from Alzheimer patients. [provided by RefSeq, Jan 2010]
UniProt Code:Q07954
NCBI GenInfo Identifier:317373384
NCBI Gene ID:4035
NCBI Accession:Q07954.2
UniProt Secondary Accession:Q07954,Q2PP12, Q86SW0, Q8IVG8,
UniProt Related Accession:Q07954
Molecular Weight:
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:APR; LRP; A2MR; CD91; APOER; LRP1A; TGFBR5; IGFBP3R  
NCBI Protein Information:prolow-density lipoprotein receptor-related protein 1; LRP-1; type V tgf-beta receptor; apolipoprotein E receptor; alpha-2-macroglobulin receptor; TbetaR-V/LRP-1/IGFBP-3 receptor
UniProt Protein Name:Prolow-density lipoprotein receptor-related protein 1
UniProt Synonym Protein Names:Alpha-2-macroglobulin receptor; A2MR; Apolipoprotein E receptor; APOER; CD_antigen: CD91Cleaved into the following 3 chains:Low-density lipoprotein receptor-related protein 1 85 kDa subunit; LRP-85; Low-density lipoprotein receptor-related protein 1 515 kDa subunit; LRP-515; Low-density lipoprotein receptor-related protein 1 intracellular domain; LRPICD
Protein Family:Low-density lipoprotein receptor-related protein
UniProt Gene Name:LRP1  
UniProt Entry Name:LRP1_HUMAN
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.
ELISA
Human LRP1 ELISA Kit

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