Human RXRa(Retinoid X Receptor Alpha)ELISA Kit (HUES03602)
- SKU:
- HUES03602
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P19793
- Sensitivity:
- 0.19ng/mL
- Range:
- 0.31-20ng/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Tested Sample Types:
- Serum, plasma and other biological fluids
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 0.31-20 ng/mL |
| Sensitivity: | 0.19 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human RXRa in samples. No significant cross-reactivity or interference between Human RXRa and analogues was observed. |
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 Human RXRa. 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 Human RXRa 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 RXRa, 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 Human RXRa. The concentration of Human RXRa in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | RXRA: Receptor for retinoic acid. Retinoic acid receptors bind as heterodimers to their target response elements in response to their ligands, all-trans or 9-cis retinoic acid, and regulate gene expression in various biological processes. The RAR/RXR heterodimers bind to the retinoic acid response elements (RARE) composed of tandem 5'-AGGTCA-3' sites known as DR1-DR5. The high affinity ligand for RXRs is 9-cis retinoic acid. RXRA serves as a common heterodimeric partner for a number of nuclear receptors. The RXR/RAR heterodimers bind to the retinoic acid response elements (RARE) composed of tandem 5'-AGGTCA-3' sites known as DR1-DR5. In the absence of ligand, the RXR-RAR heterodimers associate with a multiprotein complex containing transcription corepressors that induce histone acetylation, chromatin condensation and transcriptional suppression. On ligand binding, the corepressors dissociate from the receptors and associate with the coactivators leading to transcriptional activation. The RXRA/PPARA heterodimer is required for PPARA transcriptional activity on fatty acid oxidation genes such as ACOX1 and the P450 system genes. Homodimer. Heterodimer with RARA; required for ligand- dependent retinoic acid receptor transcriptional activity. Heterodimer with PPARA (via the leucine-like zipper in the LBD); the interaction is required for PPARA transcriptional activity. Also heterodimerizes with PPARG. Interacts with NCOA3 and NCOA6 coactivators. Interacts with FAM120B. Interacts with PELP1, SENP6, SFPQ, DNTTIP2 and RNF8. Interacts (via the DNA binding domain) with HCV core protein; the interaction enhances the transcriptional activities of the RXRA/RARA and the RXRA/PPARA heterodimers. Interacts with PRMT2. Interacts with ASXL1 and NCOA1. Highly expressed in liver, also found in lung, kidney and heart. Belongs to the nuclear hormone receptor family. NR2 subfamily. |
| UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; DNA-binding; Nuclear receptor Chromosomal Location of Human Ortholog: 9q34. 3 Cellular Component: nucleoplasm; nuclear chromatin; nucleus; receptor complex Molecular Function:ligand-dependent nuclear receptor activity; zinc ion binding; chromatin DNA binding; transcription coactivator activity; protein binding; enzyme binding; DNA binding; vitamin D receptor binding; retinoid-X receptor activity; protein heterodimerization activity; sequence-specific DNA binding; steroid hormone receptor activity; retinoic acid receptor activity; transcription factor activity Biological Process: retinoic acid receptor signaling pathway; transcription initiation from RNA polymerase II promoter; cholesterol metabolic process; camera-type eye development; vitamin metabolic process; response to retinoic acid; in utero embryonic development; ventricular cardiac muscle cell differentiation; cellular lipid metabolic process; negative regulation of transcription from RNA polymerase II promoter; positive regulation of translational initiation by iron; protein homotetramerization; virus-host interaction; ventricular cardiac muscle morphogenesis; positive regulation of transcription from RNA polymerase II promoter; gene expression; steroid hormone mediated signaling; transmembrane transport; embryo implantation; maternal placenta development |
| NCBI Summary: | Retinoid X receptors (RXRs) and retinoic acid receptors (RARs) are nuclear receptors that mediate the biological effects of retinoids by their involvement in retinoic acid-mediated gene activation. These receptors function as transcription factors by binding as homodimers or heterodimers to specific sequences in the promoters of target genes. The protein encoded by this gene is a member of the steroid and thyroid hormone receptor superfamily of transcriptional regulators. Alternative splicing of this gene results in multiple transcript variants. [provided by RefSeq, May 2014] |
| UniProt Code: | P19793 |
| NCBI GenInfo Identifier: | 133701 |
| NCBI Gene ID: | 6256 |
| NCBI Accession: | P19793. 1 |
| UniProt Secondary Accession: | P19793,Q2NL52, Q2V504, B3KY83, |
| UniProt Related Accession: | P19793 |
| Molecular Weight: | 41,060 Da |
| NCBI Full Name: | Retinoic acid receptor RXR-alpha |
| NCBI Synonym Full Names: | retinoid X receptor, alpha |
| NCBI Official Symbol: | RXRA |
| NCBI Official Synonym Symbols: | NR2B1 |
| NCBI Protein Information: | retinoic acid receptor RXR-alpha |
| UniProt Protein Name: | Retinoic acid receptor RXR-alpha |
| UniProt Synonym Protein Names: | Nuclear receptor subfamily 2 group B member 1; Retinoid X receptor alpha |
| Protein Family: | Retinoic acid receptor |
| UniProt Gene Name: | RXRA |
| UniProt Entry Name: | RXRA_HUMAN |
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.
| Concentration (ng/mL) | O.D | Average | Corrected |
| 20 | 2.506 2.526 | 2.516 | 2.447 |
| 10 | 1.635 1.667 | 1.651 | 1.582 |
| 5 | 0.974 0.966 | 0.97 | 0.901 |
| 2.5 | 0.488 0.492 | 0.49 | 0.421 |
| 1.25 | 0.277 0.263 | 0.27 | 0.201 |
| 0.63 | 0.174 0.168 | 0.171 | 0.102 |
| 0.31 | 0.12 0.122 | 0.121 | 0.052 |
| 0 | 0.064 0.074 | 0.069 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human RXRa were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human RXRa 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 (ng/mL) | 1.03 | 2.12 | 8.77 | 0.96 | 2.00 | 9.52 |
| Standard deviation | 0.06 | 0.12 | 0.33 | 0.06 | 0.11 | 0.39 |
| C V (%) | 5.83 | 5.66 | 3.76 | 6.25 | 5.50 | 4.10 |
Recovery
The recovery of Human RXRa 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) | 91-107 | 97 |
| EDTA plasma (n=5) | 89-100 | 95 |
| Cell culture media (n=5) | 83-98 | 90 |
Linearity
Samples were spiked with high concentrations of Human RXRa 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 (%) | 94-108 | 89-104 | 96-114 |
| Average (%) | 99 | 96 | 104 | |
| 1:4 | Range (%) | 89-104 | 80-91 | 84-98 |
| Average (%) | 96 | 85 | 90 | |
| 1:8 | Range (%) | 88-101 | 85-95 | 86-97 |
| Average (%) | 95 | 90 | 91 | |
| 1:16 | Range (%) | 87-99 | 84-96 | 84-94 |
| Average (%) | 93 | 89 | 89 |
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 ELISA 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 | 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 |
- 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 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl 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 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.
- 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.
- Add 100µL 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 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.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
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