Description
Rat Low-density lipoprotein receptor-related protein 2 (Lrp2) ELISA Kit
The Rat Low-Density Lipoprotein Receptor-Related Protein 2 (LRP2) ELISA Kit is a specialized assay designed for the accurate quantification of LRP2 levels in rat samples such as serum, plasma, and tissue homogenates. This kit offers high sensitivity and specificity, ensuring precise and reliable results for researchers studying the role of LRP2 in various biological processes.LRP2, also known as megalin, is a cell surface receptor that plays a critical role in the endocytosis of a wide range of ligands, including lipoproteins, hormones, and vitamins.
Its dysfunction has been implicated in various diseases, such as atherosclerosis, kidney disorders, and neurodegenerative conditions, making it a valuable biomarker for understanding these pathologies.By using the Rat LRP2 ELISA Kit, researchers can gain valuable insights into the role of this receptor in cellular processes and disease states, ultimately leading to a better understanding of its therapeutic potential. Trust in the reliability and accuracy of this kit for your research needs.
| Product Name: | Rat Low-density lipoprotein receptor-related protein 2 (Lrp2) ELISA Kit |
| SKU: | RTEB0001 |
| Size: | 96T |
| Target: | Rat Low-density lipoprotein receptor-related protein 2 (Lrp2) |
| Synonyms: | Glycoprotein 330, Megalin, gp330, LRP-2 |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Rat |
| Detection Range: | 0.625-40ng/mL |
| Sensitivity: | 0.312ng/mL |
| Intra CV: | 6.2% | ||||||||||||||||||||
| Inter CV: | 8.1% | ||||||||||||||||||||
| Linearity: |
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| Recovery: |
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| Function: | Acts together with cubilin to mediate HDL endocytosis (By similarity). Receptor-mediated uptake of polybasic drugs such as aprotinin, aminoglycosides and polymyxin B. |
| Uniprot: | P98158 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant rat Low-density lipoprotein receptor-related protein 2 |
| Sub Unit: | Binds plasminogen, extracellular matrix components, plasminogen activator-plasminogen activator inhibitor type I complex, apolipoprotein E-enriched beta-VLDL, lipoprotein lipase, lactoferrin, CLU/clusterin and calcium. Forms a multimeric complex together with LRPAP1 (PubMed:1400426). Interacts (via PxLPxI/L motif) with ANKRA2 (via ankyrin repeats) (PubMed:22649097). Interacts with LRP2BP. Interacts (via NPXY motif) with DAB2; the interaction is not affected by tyrosine phosphorylation of the NPXY motif. |
| Research Area: | Cardiovascular |
| Subcellular Location: | Cell membrane Single-pass type I membrane protein Membrane Clathrin-coated pit Cytoplasm Expressed in clathrin-coated pits, a soluble form is possibly derived by cleavage at the cell surface. |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | LRP2: Acts together with cubilin to mediate HDL endocytosis. May participate in regulation of parathyroid- hormone and para-thyroid-hormone-related protein release. Defects in LRP2 are the cause of Donnai-Barrow syndrome (DBS); also known as faciooculoacousticorenal syndrome (FOAR syndrome). DBS is a rare autosomal recessive disorder characterized by major malformations including agenesis of the corpus callosum, congenital diaphragmatic hernia, facial dysmorphology, ocular anomalies, sensorineural hearing loss and developmental delay. The FOAR syndrome was first described as comprising facial anomalies, ocular anomalies, sensorineural hearing loss, and proteinuria. DBS and FOAR were first described as distinct disorders but the classic distinguishing features between the 2 disorders were presence of proteinuria and absence of diaphragmatic hernia and corpus callosum anomalies in FOAR. Early reports noted that the 2 disorders shared many phenotypic features and may be identical. Although there is variability in the expression of some features (e.g. agenesis of the corpus callosum and proteinuria), DBS and FOAR are now considered to represent the same entity. Belongs to the LDLR family. |
| UniProt Protein Details: | Protein type:Membrane protein, integral; Receptor, misc.; Motility/polarity/chemotaxis Cellular Component: apical part of cell; apical plasma membrane; brush border; brush border membrane; coated pit; cytoplasm; endocytic vesicle; endoplasmic reticulum; endosome; extracellular space; Golgi apparatus; integral to membrane; lipid raft; lysosomal membrane; membrane; protein complex; receptor complex Molecular Function:calcium ion binding; hemoglobin binding; hormone binding; lipoprotein transporter activity; low-density lipoprotein receptor binding; PDZ domain binding; protein binding; protein complex binding; receptor activity; SH3 domain binding Biological Process: aging; cell proliferation; endosome transport; forebrain development; heart development; hemoglobin import; hormone secretion; lipoprotein transport; organ regeneration; receptor-mediated endocytosis; response to drug; response to retinoic acid; response to vitamin D; response to X-ray; transcytosis; vitamin metabolic process |
| NCBI Summary: | receptor for low density lipoprotein and a major kidney glomerular antigen for rat Heymann nephritis [RGD, Feb 2006] |
| UniProt Code: | P98158 |
| NCBI GenInfo Identifier: | 13562118 |
| NCBI Gene ID: | 29216 |
| NCBI Accession: | NP_110454.1 |
| UniProt Related Accession: | P98158 |
| Molecular Weight: | 519,276 Da |
| NCBI Full Name: | low-density lipoprotein receptor-related protein 2 |
| NCBI Synonym Full Names: | LDL receptor related protein 2 |
| NCBI Official Symbol: | Lrp2 |
| NCBI Protein Information: | low-density lipoprotein receptor-related protein 2 |
| UniProt Protein Name: | Low-density lipoprotein receptor-related protein 2 |
| UniProt Synonym Protein Names: | Glycoprotein 330; gp330; Megalin |
| Protein Family: | Low-density lipoprotein receptor-related protein |
| UniProt Gene Name: | Lrp2 |
| UniProt Entry Name: | LRP2_RAT |
| 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. |
