Human Cell Biology ELISA Kits 4
Human S100A11 (S100 Calcium Binding Protein A11) CLIA Kit (HUES00744)
- SKU:
- HUES00744
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 37.5pg/mL
- Range:
- 62.5-4000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- MLN70, S100C, Calgizzarin
- 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 S100A11 in samples. No significant cross-reactivity or interference between Human S100A11 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 S100A11. 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 S100A11 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 S100A11, 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 S100A11. The concentration of Human S100A11 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | S100A11: a calcium-binding regulatory protein of the S-100 family. Interacts with the N-termini of annexins A1. May regulate fusion processes such as endo- and exocytosis. Increases in the early stage of pancreatic carcinogenesis and decreases during subsequent progression to cancer. Increased expression in anaplastic large cell lymphoma. |
| UniProt Protein Details: | Protein type:Calcium-binding; DNA replication; Cell cycle regulation Chromosomal Location of Human Ortholog: 1q21 Cellular Component: cytoplasm; extracellular space; nucleus; ruffle Molecular Function:calcium-dependent protein binding; protein binding; protein homodimerization activity Biological Process: negative regulation of cell proliferation; negative regulation of DNA replication; signal transduction |
| NCBI Summary: | The protein encoded by this gene is a member of the S100 family of proteins containing 2 EF-hand calcium-binding motifs. S100 proteins are localized in the cytoplasm and/or nucleus of a wide range of cells, and involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. S100 genes include at least 13 members which are located as a cluster on chromosome 1q21. This protein may function in motility, invasion, and tubulin polymerization. Chromosomal rearrangements and altered expression of this gene have been implicated in tumor metastasis. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P31949 |
| NCBI GenInfo Identifier: | 1710818 |
| NCBI Gene ID: | 6282 |
| NCBI Accession: | P31949. 2 |
| UniProt Secondary Accession: | P31949,Q5VTK0, |
| UniProt Related Accession: | P31949 |
| Molecular Weight: | 11,740 Da |
| NCBI Full Name: | Protein S100-A11 |
| NCBI Synonym Full Names: | S100 calcium binding protein A11 |
| NCBI Official Symbol: | S100A11 |
| NCBI Official Synonym Symbols: | MLN70; S100C; HEL-S-43 |
| NCBI Protein Information: | protein S100-A11 |
| UniProt Protein Name: | Protein S100-A11 |
| UniProt Synonym Protein Names: | Calgizzarin; Metastatic lymph node gene 70 protein; MLN 70 |
| Protein Family: | Protein |
| UniProt Gene Name: | S100A11 |
| UniProt Entry Name: | S10AB_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 | 53808 64354 | 59081 | 59046 |
| 2000 | 27686 28032 | 27859 | 27824 |
| 1000 | 13975 12993 | 13484 | 13449 |
| 500 | 6187 7023 | 6605 | 6570 |
| 250 | 3319 3167 | 3243 | 3208 |
| 125 | 1638 1524 | 1581 | 1546 |
| 62.50 | 743 767 | 755 | 720 |
| 0 | 35 35 | 35 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human S100A11 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human S100A11 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) | 218.13 | 537.35 | 1645.33 | 238.26 | 510.96 | 1712.52 |
| Standard deviation | 21.97 | 56.80 | 148.41 | 27.23 | 40.06 | 132.72 |
| C V (%) | 10.07 | 10.57 | 9.02 | 11.43 | 7.84 | 7.75 |
Recovery
The recovery of Human S100A11 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) | 96-109 | 102 |
| EDTA plasma (n=5) | 91-106 | 99 |
| Cell culture media (n=5) | 93-107 | 101 |
Linearity
Samples were spiked with high concentrations of Human S100A11 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 (%) | 99-115 | 85-97 | 85-99 |
| Average (%) | 105 | 92 | 91 | |
| 1:4 | Range (%) | 95-108 | 94-107 | 93-103 |
| Average (%) | 101 | 101 | 98 | |
| 1:8 | Range (%) | 93-109 | 86-102 | 94-108 |
| Average (%) | 99 | 93 | 102 | |
| 1:16 | Range (%) | 89-103 | 101-117 | 86-100 |
| Average (%) | 96 | 107 | 92 |
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.
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Human Cell Biology ELISA Kits 6
