Human Cell Biology ELISA Kits 3
Human CHEM (Chemerin) CLIA Kit (HUES00448)
- SKU:
- HUES00448
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 18.75pg/mL
- Range:
- 31.25-2000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- RARRES2, TIG2, Retinoic acid receptor responder protein 2, Tazarotene-induced gene 2 protein
- 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: | 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 Human CHEM in samples. No significant cross-reactivity or interference between Human CHEM 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 CHEM. 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 CHEM 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 CHEM, 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 CHEM. The concentration of Human CHEM in the samples can be calculated by comparing the RLU of the samples to the standard curve.
UniProt Protein Function: | RARRES2: Inhibited in psoriatic lesions. Activated by tazarotene in skin rafts and in the epidermis of psoriatic lesions. |
UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide Chromosomal Location of Human Ortholog: 7q36. 1 Cellular Component: extracellular matrix; extracellular region Molecular Function:protein binding; receptor binding Biological Process: cell differentiation; chemotaxis; embryonic gut development; in utero embryonic development; inflammatory response; positive regulation of chemotaxis; positive regulation of fat cell differentiation; positive regulation of protein amino acid phosphorylation; regulation of lipid catabolic process; retinoid metabolic process |
NCBI Summary: | This gene encodes a secreted chemotactic protein that initiates chemotaxis via the ChemR23 G protein-coupled seven-transmembrane domain ligand. Expression of this gene is upregulated by the synthetic retinoid tazarotene and occurs in a wide variety of tissues. The active protein has several roles, including that as an adipokine and as an antimicrobial protein with activity against bacteria and fungi. [provided by RefSeq, Nov 2014] |
UniProt Code: | Q99969 |
NCBI GenInfo Identifier: | 6226227 |
NCBI Gene ID: | 5919 |
NCBI Accession: | Q99969. 1 |
UniProt Secondary Accession: | Q99969,Q7LE02, |
UniProt Related Accession: | Q99969 |
Molecular Weight: | 18,618 Da |
NCBI Full Name: | Retinoic acid receptor responder protein 2 |
NCBI Synonym Full Names: | retinoic acid receptor responder 2 |
NCBI Official Symbol: | RARRES2 |
NCBI Official Synonym Symbols: | TIG2; HP10433 |
NCBI Protein Information: | retinoic acid receptor responder protein 2 |
UniProt Protein Name: | Retinoic acid receptor responder protein 2 |
UniProt Synonym Protein Names: | Chemerin; RAR-responsive protein TIG2; Tazarotene-induced gene 2 protein |
Protein Family: | Retinoic acid receptor responder protein |
UniProt Gene Name: | RARRES2 |
UniProt Entry Name: | RARR2_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 |
2000 | 53881 57561 | 55721 | 55693 |
1000 | 22881 25067 | 23974 | 23946 |
500 | 11524 10540 | 11032 | 11004 |
250 | 4882 5706 | 5294 | 5266 |
125 | 2815 2401 | 2608 | 2580 |
62.5 | 1387 1235 | 1311 | 1283 |
31.25 | 662 686 | 674 | 646 |
0 | 28 28 | 28 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CHEM were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CHEM 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) | 90.96 | 236.44 | 670.99 | 94.29 | 243.16 | 604.29 |
Standard deviation | 8.88 | 17.78 | 69.18 | 9.70 | 22.44 | 51.06 |
C V (%) | 9.76 | 7.52 | 10.31 | 10.29 | 9.23 | 8.45 |
Recovery
The recovery of Human CHEM 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) | 91-103 | 96 |
EDTA plasma (n=5) | 94-109 | 101 |
Cell culture media (n=5) | 95-107 | 102 |
Linearity
Samples were spiked with high concentrations of Human CHEM 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 (%) | 89-104 | 85-97 | 92-109 |
Average (%) | 95 | 91 | 100 | |
1:4 | Range (%) | 97-114 | 89-103 | 91-104 |
Average (%) | 104 | 94 | 97 | |
1:8 | Range (%) | 88-99 | 103-117 | 96-106 |
Average (%) | 93 | 109 | 101 | |
1:16 | Range (%) | 100-116 | 98-110 | 100-118 |
Average (%) | 108 | 104 | 108 |
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 100õl of standard solutions into the standard wells.
- Add 100õl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100õl 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 37ðC.
- Aspirate the liquid from each well, do not wash. Immediately add 100õL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37ðC.
- Aspirate or decant the solution from the plate and add 350õL 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 100õL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37ðC.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100õL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37ðC. Protect the plate from light.
- Determine the RLU value of each well immediately.