The Mouse Endothelin B Receptor (EDNRB) ELISA Kit is a powerful tool for the precise measurement of EDNRB levels in mouse samples. With exceptional sensitivity and specificity, this kit delivers consistent and accurate results, making it invaluable for a variety of research endeavors.EDNRB is a key receptor involved in endothelin signaling pathways, influencing various physiological processes such as vascular tone regulation and cell growth. Dysregulation of EDNRB activity has been linked to several diseases including pulmonary hypertension, cancer, and inflammatory conditions, highlighting its significance as a potential therapeutic target.
This ELISA kit provides researchers with a reliable means to study EDNRB expression levels in biological samples, aiding in the understanding of its role in normal physiology and disease pathogenesis. Trusted by scientists worldwide, this kit is an essential asset for studies aiming to unravel the complexities of endothelin signaling and its implications in health and disease.
Product Name:
Mouse Endothelin B receptor (Ednrb) ELISA Kit
SKU:
MOEB1830
Size:
96T
Target:
Mouse Endothelin B receptor (Ednrb)
Synonyms:
Endothelin receptor non-selective type, ET-B
Assay Type:
Sandwich
Detection Method:
ELISA
Reactivity:
Mouse
Detection Range:
78-5000pg/mL
Sensitivity:
39pg/mL
Intra CV:
6.4%
Inter CV:
9.0%
Linearity:
Sample
1:2
1:4
1:8
1:16
Serum(N=5)
112-112%
110-119%
114-126%
109-119%
EDTA Plasma(N=5)
100-110%
102-112%
106-116%
110-118%
Heparin Plasma(N=5)
101-111%
88-99%
82-90%
94-106%
Recovery:
Sample Type
Average(%)
Recovery Range(%)
Serum
99
93-105
Plasma
101
95-107
Function:
Non-specific receptor for endothelin 1, 2, and 3. Mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system. Essential component in the normal development of two neuronal crest-derived cell lineages.
Uniprot:
P48302
Sample Type:
Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:
Natural and recombinant mouse Endothelin receptor type B
Subcellular Location:
Cell membrane Multi-pass membrane protein internalized after activation by endothelins.
Storage:
Please see kit components below for exact storage details
Note:
For research use only
UniProt Protein Function:
ETB: a G protein-coupled receptor which activates a phosphatidylinoitol-calcium second messenger system. Its ligand, endothelin, consists of a family of three potent vasoactive peptides: ET1, ET2, and ET3. Studies suggest that the multigenic disorder, Hirschsprung disease type 2, is due to mutation in endothelin receptor type B gene. Essential component in the normal development of two neuronal crest-derived cell lineages. Two splice variant isoforms have been described.Protein type: Membrane protein, integral; GPCR, family 1; Receptor, GPCR; Membrane protein, multi-passChromosomal Location of Human Ortholog: 13q22Cellular Component: nuclear membrane; integral to plasma membrane; plasma membrane; lipid raftMolecular Function: endothelin receptor activity; protein binding; peptide hormone binding; type 1 angiotensin receptor bindingBiological Process: epithelial fluid transport; cGMP-mediated signaling; regulation of fever; response to pain; negative regulation of cellular protein metabolic process; negative regulation of transcription from RNA polymerase II promoter; vein smooth muscle contraction; sensory perception of pain; vasodilation; response to organic cyclic substance; enteric nervous system development; regulation of epithelial cell proliferation; elevation of cytosolic calcium ion concentration; cell surface receptor linked signal transduction; regulation of blood pressure; positive regulation of cell proliferation; melanocyte differentiation; regulation of pH; neural crest cell migration; aging; nervous system development; posterior midgut development; vasoconstriction; negative regulation of adenylate cyclase activity; macrophage chemotaxis; peripheral nervous system development; negative regulation of neuron maturation; G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); regulation of sensory perception of pain; positive regulation of protein amino acid phosphorylation; negative regulation of apoptosisDisease: Abcd Syndrome; Waardenburg Syndrome, Type 4a; Hirschsprung Disease, Susceptibility To, 2
UniProt Protein Details:
NCBI Summary:
The protein encoded by this gene is a G protein-coupled receptor which activates a phosphatidylinositol-calcium second messenger system. Its ligand, endothelin, consists of a family of three potent vasoactive peptides: ET1, ET2, and ET3. Studies suggest that the multigenic disorder, Hirschsprung disease type 2, is due to mutations in the endothelin receptor type B gene. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jan 2011]
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.
