|Detection range:||31.25-2000 pg/mL|
|Sample type:||Serum, plasma and other biological fluids|
|Repeatability:||CV < 15%|
|Specificity:||This kit recognizes Human NUMA1 in samples. No significant cross-reactivity or interference between Human NUMA1 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 NUMA1. 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 NUMA1 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 NUMA1, 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 NUMA1. The concentration of Human NUMA1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
|UniProt Protein Function:||NuMA-1: a coiled-coil nuclear protein that is required to establish and maintain mammalian spindle poles. Dissociates from condensing chromosomes during early prophase, before the complete disintegration of the nuclear lamina. As mitosis progresses it reassociates with telophase chromosomes very early during nuclear reformation, before substantial accumulation of lamins on chromosomal surfaces is evident. Two human splice-variant isoforms have been described.|
|UniProt Protein Details:|
Protein type:Cell cycle regulation; Motility/polarity/chemotaxis
Chromosomal Location of Human Ortholog: 11q13
Cellular Component: spindle pole; nuclear matrix; dendrite; chromosome; cytosol; nucleoplasm; Golgi membrane; cell soma; apical part of cell; spindle microtubule; cytoplasm; spindle; nucleus
Molecular Function:protein binding; microtubule binding; structural molecule activity
Biological Process: mitosis; meiotic cell cycle; establishment of mitotic spindle orientation; cell division; mitotic cell cycle; G2/M transition of mitotic cell cycle; nuclear organization and biogenesis
Disease: Acute Promyelocytic Leukemia
|NCBI Summary:||This gene encodes a large protein that forms a structural component of the nuclear matrix. The encoded protein interacts with microtubules and plays a role in the formation and organization of the mitotic spindle during cell division. Chromosomal translocation of this gene with the RARA (retinoic acid receptor, alpha) gene on chromosome 17 have been detected in patients with acute promyelocytic leukemia. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Nov 2013]|
|NCBI GenInfo Identifier:||145559510|
|NCBI Gene ID:||4926|
|NCBI Accession:||Q14980. 2|
|UniProt Secondary Accession:||Q14980,Q14981, Q9BTE9, H0YH75,|
|UniProt Related Accession:||Q14980|
|Molecular Weight:||109,279 Da|
|NCBI Full Name:||Nuclear mitotic apparatus protein 1|
|NCBI Synonym Full Names:||nuclear mitotic apparatus protein 1|
|NCBI Official Symbol:||NUMA1|
|NCBI Official Synonym Symbols:||NUMA|
|NCBI Protein Information:||nuclear mitotic apparatus protein 1; SP-H antigen; structural nuclear protein; centrophilin stabilizes mitotic spindle in mitotic cells|
|UniProt Protein Name:||Nuclear mitotic apparatus protein 1|
|UniProt Synonym Protein Names:||Nuclear matrix protein-22|
|Protein Family:||Nuclear mitotic apparatus protein|
|UniProt Gene Name:||NUMA1|
|UniProt Entry Name:||NUMA1_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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human NUMA1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NUMA1 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||12.20||11.78||11.49||8.49||8.51||7.54|
The recovery of Human NUMA1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||91-104||97|
|Cell culture media (n=5)||95-110||101|
Samples were spiked with high concentrations of Human NUMA1 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)|
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.
|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.