Human Cell Biology ELISA Kits 5
Human ETRB (Endothelin Receptor B) CLIA Kit (HUES00992)
- SKU:
- HUES00992
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 37.5pg/mL
- Range:
- 62.5-4000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- EDNRB, ETB, HSCR, HSCR2
- 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: | Chemiluminescence |
Detection range: | 62.50-4000 pg/mL |
Sensitivity: | 37.50 pg/mL |
Sample volume: | 100µL |
Sample type: | Serum, plasma and other biological fluids |
Repeatability: | CV < 15% |
Specificity: | This kit recognizes Human ETRB in samples. No significant cross-reactivity or interference between Human ETRB and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human ETRB. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human ETRB 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 ETRB, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human ETRB. The concentration of Human ETRB in the samples can be calculated by comparing the RLU of the samples to the standard curve.
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. |
UniProt Protein Details: | Protein type:Membrane protein, integral; GPCR, family 1; Membrane protein, multi-pass; Receptor, GPCR Chromosomal Location of Human Ortholog: 13q22 Cellular Component: nuclear membrane; integral to plasma membrane; plasma membrane; lipid raft Molecular Function:endothelin receptor activity; protein binding; peptide hormone binding; type 1 angiotensin receptor binding Biological Process: epithelial fluid transport; cGMP-mediated signaling; regulation of fever; negative regulation of cellular protein metabolic process; response to pain; negative regulation of transcription from RNA polymerase II promoter; vein smooth muscle contraction; sensory perception of pain; response to organic cyclic substance; enteric nervous system development; vasodilation; regulation of epithelial cell proliferation; elevation of cytosolic calcium ion concentration; cell surface receptor linked signal transduction; regulation of blood pressure; melanocyte differentiation; positive regulation of cell proliferation; regulation of pH; neural crest cell migration; aging; posterior midgut development; nervous system 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); positive regulation of protein amino acid phosphorylation; regulation of sensory perception of pain; negative regulation of apoptosis Disease: Waardenburg Syndrome, Type 4a; Abcd Syndrome; Hirschsprung Disease, Susceptibility To, 2 |
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] |
UniProt Code: | P24530 |
NCBI GenInfo Identifier: | 119622 |
NCBI Gene ID: | 1910 |
NCBI Accession: | P24530. 1 |
UniProt Secondary Accession: | P24530,O15343, Q59GB1, Q5W0G9, Q8NHM6, Q8NHM7, Q8NHM8 Q8NHM9, Q9UD23, Q9UQK3, A2A2Z8, A8K3T4, |
UniProt Related Accession: | P24530,AAB24922 |
Molecular Weight: | 436 |
NCBI Full Name: | Endothelin B receptor |
NCBI Synonym Full Names: | endothelin receptor type B |
NCBI Official Symbol: | EDNRB |
NCBI Official Synonym Symbols: | ETB; ET-B; ETBR; ETRB; HSCR; WS4A; ABCDS; ET-BR; HSCR2 |
NCBI Protein Information: | endothelin B receptor; endothelin receptor non-selective type |
UniProt Protein Name: | Endothelin B receptor |
UniProt Synonym Protein Names: | Endothelin receptor non-selective type |
Protein Family: | Endothelin B receptor |
UniProt Gene Name: | EDNRB |
UniProt Entry Name: | EDNRB_HUMAN |
As the RLU 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 (pg/mL) | RLU | Average | Corrected |
4000 | 54306 63856 | 59081 | 59046 |
2000 | 26293 29425 | 27859 | 27824 |
1000 | 14677 12291 | 13484 | 13449 |
500 | 6518 6692 | 6605 | 6570 |
250 | 3280 3206 | 3243 | 3208 |
125 | 1621 1541 | 1581 | 1546 |
62.50 | 737 773 | 755 | 720 |
0 | 34 36 | 35 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human ETRB were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human ETRB 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 (pg/mL) | 189.48 | 469.11 | 1836.27 | 198.84 | 457.77 | 1768.73 |
Standard deviation | 18.85 | 40.91 | 198.50 | 22.01 | 46.51 | 159.01 |
C V (%) | 9.95 | 8.72 | 10.81 | 11.07 | 10.16 | 8.99 |
Recovery
The recovery of Human ETRB 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) | 91-105 | 97 |
EDTA plasma (n=5) | 98-110 | 104 |
Cell culture media (n=5) | 91-106 | 97 |
Linearity
Samples were spiked with high concentrations of Human ETRB 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 (%) | 99-117 | 84-96 | 88-99 |
Average (%) | 106 | 91 | 93 | |
1:4 | Range (%) | 102-117 | 94-106 | 95-109 |
Average (%) | 109 | 101 | 101 | |
1:8 | Range (%) | 86-96 | 93-107 | 101-113 |
Average (%) | 91 | 100 | 107 | |
1:16 | Range (%) | 87-103 | 100-118 | 100-115 |
Average (%) | 94 | 108 | 107 |
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 CLIA 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 A | 1 vial, 5 mL | 4°C (shading light) |
Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
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 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL 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 100 µL 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 350 µL 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 100 µL 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 100 µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37 °C. Protect the plate from light.
- Determine the RLU value of each well immediately.