Rat Signaling ELISA Kits 3
Rat TrxR1 (Thioredoxin Reductase 1) CLIA Kit (RTES00542)
- SKU:
- RTES00542
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 18.75pg/mL
- Range:
- 31.25-2000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- TXNRD1, GRIM-12, TR, TR1, TXNR
- Reactivity:
- Rat
- Sample Type:
- Serum, plasma and other biological fluids
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Rat |
| Detection method: | Chemiluminescence |
| Detection range: | 31.25-2000 pg/mL |
| Sensitivity: | 18.75 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Rat TrxR1 in samples. No significant cross-reactivity or interference between Rat TrxR1 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 Rat TrxR1. 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 Rat TrxR1 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 Rat TrxR1, 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 Rat TrxR1. The concentration of Rat TrxR1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | TRXR1: a cytoplasmic pyridine nucleotide oxidoreductases. This protein reduces thioredoxins as well as other substrates, and plays a role in selenium metabolism and protection against oxidative stress. The functional enzyme is thought to be a homodimer which uses FAD as a cofactor. Each subunit contains a selenocysteine residue which is required for the catalytic activity. |
| UniProt Protein Details: | Protein type:EC 1. 8. 1. 9; Nuclear receptor co-regulator; Nucleotide Metabolism - pyrimidine; Oxidoreductase Chromosomal Location of Human Ortholog: 7q13 Cellular Component: cell; cell soma; cytoplasm; cytosol; fibrillar center; mitochondrion; nucleoplasm; nucleus Molecular Function:electron transfer activity; FAD binding; mercury ion binding; NAD(P)H oxidase activity; protein homodimerization activity; selenate reductase activity; thioredoxin-disulfide reductase activity Biological Process: benzene-containing compound metabolic process; cell proliferation; cell redox homeostasis; gastrulation; hydrogen peroxide catabolic process; mesoderm formation; placenta development; protein tetramerization; response to axon injury; response to drug; response to hyperoxia; response to oxidative stress; response to oxygen radical; response to selenium ion; selenocysteine metabolic process |
| NCBI Summary: | The protein encoded by this gene belongs to the pyridine nucleotide-disulfide oxidoreductase family, and is a member of the thioredoxin (Trx) system. Three thioredoxin reductase (TrxR) isozymes are found in mammals. TrxRs are selenocysteine-containing flavoenzymes, which reduce thioredoxins, as well as other substrates, and play a key role in redox homoeostasis. This gene encodes an ubiquitously expressed, cytosolic form of TrxR, which functions as a homodimer containing FAD, and selenocysteine (Sec) at the active site. Sec is encoded by UGA codon that normally signals translation termination. The 3' UTRs of selenoprotein mRNAs contain a conserved stem-loop structure, the Sec insertion sequence (SECIS) element, which is necessary for the recognition of UGA as a Sec codon rather than as a stop signal. Alternative splicing, primarily at the 5' end, results in transcript variants encoding same or different isoforms. [provided by RefSeq, Jun 2017] |
| UniProt Code: | O89049 |
| NCBI GenInfo Identifier: | 357529586 |
| NCBI Gene ID: | 58819 |
| NCBI Accession: | O89049. 5 |
| UniProt Secondary Accession: | O89049,Q5U344, Q9JKZ3, Q9JKZ4, Q9R1I3, |
| UniProt Related Accession: | O89049 |
| Molecular Weight: | 54,670 Da |
| NCBI Full Name: | Thioredoxin reductase 1, cytoplasmic |
| NCBI Synonym Full Names: | thioredoxin reductase 1 |
| NCBI Official Symbol: | Txnrd1 |
| NCBI Official Synonym Symbols: | Tr |
| NCBI Protein Information: | thioredoxin reductase 1, cytoplasmic |
| UniProt Protein Name: | Thioredoxin reductase 1, cytoplasmic |
| UniProt Synonym Protein Names: | NADPH-dependent thioredoxin reductase; Thioredoxin reductase TR1 |
| Protein Family: | Thioredoxin reductase |
| UniProt Gene Name: | Txnrd1 |
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 |
| 2000 | 52285 59157 | 55721 | 55693 |
| 1000 | 21591 26357 | 23974 | 23946 |
| 500 | 11599 10465 | 11032 | 11004 |
| 250 | 5206 5382 | 5294 | 5266 |
| 125 | 2732 2484 | 2608 | 2580 |
| 62.5 | 1370 1252 | 1311 | 1283 |
| 31.25 | 651 697 | 674 | 646 |
| 0 | 27 29 | 28 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat TrxR1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat TrxR1 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) | 94.87 | 312.86 | 818.22 | 103.91 | 340.67 | 786.92 |
| Standard deviation | 9.16 | 24.00 | 87.71 | 11.80 | 36.96 | 73.58 |
| C V (%) | 9.66 | 7.67 | 10.72 | 11.36 | 10.85 | 9.35 |
Recovery
The recovery of Rat TrxR1 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) | 91-108 | 98 |
| EDTA plasma (n=5) | 88-104 | 95 |
| Cell culture media (n=5) | 98-115 | 106 |
Linearity
Samples were spiked with high concentrations of Rat TrxR1 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 (%) | 87-98 | 90-104 | 92-102 |
| Average (%) | 93 | 96 | 97 | |
| 1:4 | Range (%) | 95-107 | 92-104 | 96-109 |
| Average (%) | 101 | 97 | 101 | |
| 1:8 | Range (%) | 85-100 | 102-117 | 97-113 |
| Average (%) | 91 | 109 | 103 | |
| 1:16 | Range (%) | 98-109 | 99-113 | 83-96 |
| Average (%) | 104 | 106 | 90 |
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 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 100µL 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.
Related Products
