Human Cell Biology ELISA Kits 3
Human MYH10 (Myosin Heavy Chain 10, Non Muscle) CLIA Kit (HUES00456)
- SKU:
- HUES00456
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 18.75pg/mL
- Range:
- 31.25-2000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- NMMHC-IIB, NMMHCB
- 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 MYH10 in samples. No significant cross-reactivity or interference between Human MYH10 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 MYH10. 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 MYH10 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 MYH10, 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 MYH10. The concentration of Human MYH10 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
UniProt Protein Function: | MYH10: Cellular myosin that appears to play a role in cytokinesis, cell shape, and specialized functions such as secretion and capping. Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2. Myosin is a hexameric protein that consists of 2 heavy chain subunits (MHC), 2 alkali light chain subunits (MLC) and 2 regulatory light chain subunits (MLC-2). Interacts with PLEKHG6. Interacts with ECM29. Interacts with KIF26B. Interacts with LARP6. Isoform 1 is expressed in cerebellum and spinal chord. Isoform 2 is expressed in cerebrum and retina. Isoform 3 is expressed in the cerebrum and to a much lower extent in cerebellum. 3 isoforms of the human protein are produced by alternative splicing. |
UniProt Protein Details: | Protein type:Motility/polarity/chemotaxis; Motor Chromosomal Location of Human Ortholog: 17p13 Cellular Component: mitochondrion; myosin II complex; dendritic spine; actomyosin; cell cortex; actin cytoskeleton; growth cone; cell soma; axon; lamellipodium; cytoplasm; plasma membrane; stress fiber; spindle; midbody; neuromuscular junction; nucleus; myosin complex; cleavage furrow Molecular Function:calmodulin binding; microfilament motor activity; actin filament binding; protein binding; ADP binding; actin-dependent ATPase activity; actin binding; ATP binding Biological Process: adult heart development; axon guidance; exocytosis; metabolic process; actin filament-based movement; in utero embryonic development; fourth ventricle development; ventricular cardiac muscle cell development; actomyosin structure organization and biogenesis; cerebellar Purkinje cell layer development; neuron migration; cardiac myofibril assembly; third ventricle development; substrate-bound cell migration, cell extension; regulation of cell shape; cell proliferation; cytokinesis after mitosis; plasma membrane repair; retina development in camera-type eye; ephrin receptor signaling pathway; neuromuscular process controlling balance; cell adhesion; nuclear migration; lateral ventricle development |
NCBI Summary: | This gene encodes a member of the myosin superfamily. The protein represents a conventional non-muscle myosin; it should not be confused with the unconventional myosin-10 (MYO10). Myosins are actin-dependent motor proteins with diverse functions including regulation of cytokinesis, cell motility, and cell polarity. Mutations in this gene have been associated with May-Hegglin anomaly and developmental defects in brain and heart. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Dec 2011] |
UniProt Code: | P35580 |
NCBI GenInfo Identifier: | 215274129 |
NCBI Gene ID: | 4628 |
NCBI Accession: | P35580. 3 |
UniProt Related Accession: | P35580 |
Molecular Weight: | 250kDa |
NCBI Full Name: | Myosin-10 |
NCBI Synonym Full Names: | myosin heavy chain 10 |
NCBI Official Symbol: | MYH10 |
NCBI Official Synonym Symbols: | NMMHCB; NMMHC-IIB |
NCBI Protein Information: | myosin-10 |
UniProt Protein Name: | Myosin-10 |
UniProt Synonym Protein Names: | Cellular myosin heavy chain, type B; Myosin heavy chain 10; Myosin heavy chain, non-muscle IIb; Non-muscle myosin heavy chain B; NMMHC-B; Non-muscle myosin heavy chain IIb; NMMHC II-b; NMMHC-IIB |
Protein Family: | Myosin |
UniProt Gene Name: | MYH10 |
UniProt Entry Name: | MYH10_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 | 53422 58020 | 55721 | 55693 |
1000 | 21740 26208 | 23974 | 23946 |
500 | 11578 10486 | 11032 | 11004 |
250 | 5084 5504 | 5294 | 5266 |
125 | 2657 2559 | 2608 | 2580 |
62.5 | 1322 1300 | 1311 | 1283 |
31.25 | 655 693 | 674 | 646 |
0 | 27 29 | 28 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MYH10 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MYH10 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) | 94.83 | 216.35 | 805.12 | 93.87 | 224.66 | 791.66 |
Standard deviation | 9.67 | 17.09 | 85.58 | 9.57 | 19.68 | 52.49 |
C V (%) | 10.20 | 7.90 | 10.63 | 10.19 | 8.76 | 6.63 |
Recovery
The recovery of Human MYH10 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) | 85-100 | 91 |
EDTA plasma (n=5) | 96-111 | 103 |
Cell culture media (n=5) | 94-109 | 100 |
Linearity
Samples were spiked with high concentrations of Human MYH10 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 (%) | 88-100 | 97-115 | 93-106 |
Average (%) | 95 | 105 | 99 | |
1:4 | Range (%) | 85-98 | 92-106 | 100-113 |
Average (%) | 90 | 99 | 106 | |
1:8 | Range (%) | 86-100 | 96-109 | 97-110 |
Average (%) | 93 | 102 | 104 | |
1:16 | Range (%) | 100-113 | 85-101 | 87-101 |
Average (%) | 106 | 92 | 94 |
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.