Human Cell Biology ELISA Kits 5
Human KRT10 (Keratin, Type I Cytoskeletal 10) CLIA Kit (HUES01129)
- SKU:
- HUES01129
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 4.69pg/mL
- Range:
- 7.81-500pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- CK10, K10, Cytokeratin-10, Keratin-10, EHK, KPP, BCIE, BIE
- 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: | 7.81-500 pg/mL |
| Sensitivity: | 4.69 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human KRT10 in samples. No significant cross-reactivity or interference between Human KRT10 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 KRT10. 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 KRT10 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 KRT10, 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 KRT10. The concentration of Human KRT10 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | K10: a type I cytoskeletal keratin. The keratins are intermediate filament proteins responsible for the structural integrity of epithelial cells and are subdivided into cytokeratins and hair keratins. There are two types of cytoskeletal and microfibrillar keratin: type I (acidic; 40-55 kDa) [K9 to K20] and type II (neutral to basic; 56-70 kDa) [K1 to K8]. Both a basic and an acidic keratin are required for filament assembly. Generally associates with K1. |
| UniProt Protein Details: | Protein type:Cytoskeletal Chromosomal Location of Human Ortholog: 17q21 Cellular Component: cytoplasm; extracellular space; membrane; nucleus Biological Process: keratinocyte differentiation Disease: Epidermolytic Hyperkeratosis; Erythroderma, Ichthyosiform, Congenital Reticular; Ichthyosis, Cyclic, With Epidermolytic Hyperkeratosis |
| NCBI Summary: | This gene encodes a member of the type I (acidic) cytokeratin family, which belongs to the superfamily of intermediate filament (IF) proteins. Keratins are heteropolymeric structural proteins which form the intermediate filament. These filaments, along with actin microfilaments and microtubules, compose the cytoskeleton of epithelial cells. Mutations in this gene are associated with epidermolytic hyperkeratosis. This gene is located within a cluster of keratin family members on chromosome 17q21. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P13645 |
| NCBI GenInfo Identifier: | 269849769 |
| NCBI Gene ID: | 3858 |
| NCBI Accession: | P13645. 6 |
| UniProt Secondary Accession: | P13645,Q14664, Q8N175, |
| UniProt Related Accession: | P13645 |
| Molecular Weight: | 58,827 Da |
| NCBI Full Name: | Keratin, type I cytoskeletal 10 |
| NCBI Synonym Full Names: | keratin 10 |
| NCBI Official Symbol: | KRT10 |
| NCBI Official Synonym Symbols: | BIE; EHK; K10; KPP; BCIE; CK10 |
| NCBI Protein Information: | keratin, type I cytoskeletal 10 |
| UniProt Protein Name: | Keratin, type I cytoskeletal 10 |
| UniProt Synonym Protein Names: | Cytokeratin-10; CK-10; Keratin-10; K10 |
| Protein Family: | Keratin |
| UniProt Gene Name: | KRT10 |
| UniProt Entry Name: | K1C10_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 |
| 500 | 48166 54908 | 51537 | 51508 |
| 250 | 19509 22487 | 20998 | 20969 |
| 125 | 9581 9211 | 9396 | 9367 |
| 62.5 | 4137 4887 | 4512 | 4483 |
| 31.25 | 2356 2242 | 2299 | 2270 |
| 15.63 | 1264 1236 | 1250 | 1221 |
| 7.81 | 691 789 | 740 | 711 |
| 0 | 28 30 | 29 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human KRT10 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human KRT10 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) | 27.48 | 65.37 | 171.25 | 27.44 | 67.27 | 165.60 |
| Standard deviation | 2.70 | 6.92 | 10.28 | 3.36 | 5.11 | 12.04 |
| C V (%) | 9.83 | 10.59 | 6.00 | 12.24 | 7.60 | 7.27 |
Recovery
The recovery of Human KRT10 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) | 97-108 | 102 |
| EDTA plasma (n=5) | 88-103 | 96 |
| Cell culture media (n=5) | 86-99 | 91 |
Linearity
Samples were spiked with high concentrations of Human KRT10 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 (%) | 100-113 | 94-110 | 88-102 |
| Average (%) | 107 | 101 | 95 | |
| 1:4 | Range (%) | 101-113 | 86-99 | 96-111 |
| Average (%) | 107 | 91 | 102 | |
| 1:8 | Range (%) | 88-102 | 84-96 | 96-113 |
| Average (%) | 93 | 90 | 104 | |
| 1:16 | Range (%) | 90-103 | 97-111 | 98-115 |
| Average (%) | 97 | 104 | 105 |
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.
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