Human CCNB (Cyclin-B) CLIA Kit (HUES00214)
- Product type:
- ELISA Kit
- ELISA Type:
|Detection range:||78.13-5000 pg/mL|
|Sample type:||Serum, plasma and other biological fluids|
|Repeatability:||CV < 15%|
|Specificity:||This kit recognizes Human CCNB in samples. No significant cross-reactivity or interference between Human CCNB 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 CCNB. 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 CCNB 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 CCNB, 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 CCNB. The concentration of Human CCNB in the samples can be calculated by comparing the RLU of the samples to the standard curve.
|UniProt Protein Function:||CCNB1: a member of the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance through the cell cycle. Cyclins function as regulators of CDK kinases. Complexes with cdc2 to form the maturation-promoting factor (MPF). Two alternatively spliced isoforms have been found, a constitutively expressed form and a cell cycle-regulated form that is expressed predominantly during G2/M phase.|
|UniProt Protein Details:|
Protein type:Cell cycle regulation; Activator
Chromosomal Location of Human Ortholog: 5q12
Cellular Component: nucleoplasm; spindle pole; centrosome; membrane; cytoplasm; nucleus; cytosol
Molecular Function:protein binding; patched binding; histone kinase activity; protein kinase binding
Biological Process: positive regulation of ubiquitin-protein ligase activity during mitotic cell cycle; positive regulation of histone phosphorylation; regulation of cell cycle; ventricular cardiac muscle cell development; positive regulation of attachment of spindle microtubules to kinetochore; mitotic nuclear envelope disassembly; positive regulation of mRNA 3'-end processing; positive regulation of mitotic cell cycle; positive regulation of fibroblast proliferation; negative regulation of protein amino acid phosphorylation; tissue regeneration; anaphase-promoting complex-dependent proteasomal ubiquitin-dependent protein catabolic process; protein complex assembly; G2/M transition of mitotic cell cycle; positive regulation of cardiac muscle cell proliferation; response to drug; in utero embryonic development; oocyte maturation; mitotic metaphase plate congression; response to DDT; gut development; regulation of ubiquitin-protein ligase activity during mitotic cell cycle; response to mechanical stimulus; cell division; spermatogenesis; regulation of cyclin-dependent protein kinase activity; mitotic cell cycle; mitotic spindle stabilization; G1/S transition of mitotic cell cycle
|NCBI Summary:||The protein encoded by this gene is a regulatory protein involved in mitosis. The gene product complexes with p34(cdc2) to form the maturation-promoting factor (MPF). Two alternative transcripts have been found, a constitutively expressed transcript and a cell cycle-regulated transcript, that is expressed predominantly during G2/M phase. The different transcripts result from the use of alternate transcription initiation sites. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||116176|
|NCBI Gene ID:||891|
|NCBI Accession:||P14635. 1|
|UniProt Secondary Accession:||P14635,Q5TZP9, A8K066,|
|UniProt Related Accession:||P14635|
|Molecular Weight:||44,102 Da|
|NCBI Full Name:||G2/mitotic-specific cyclin-B1|
|NCBI Synonym Full Names:||cyclin B1|
|NCBI Official Symbol:||CCNB1|
|NCBI Official Synonym Symbols:||CCNB|
|NCBI Protein Information:||G2/mitotic-specific cyclin-B1; G2/mitotic-specific cyclin B1|
|UniProt Protein Name:||G2/mitotic-specific cyclin-B1|
|UniProt Gene Name:||CCNB1|
|UniProt Entry Name:||CCNB1_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 CCNB were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CCNB were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||12.90||10.98||8.50||10.37||8.51||6.30|
The recovery of Human CCNB 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)||86-99||91|
|Cell culture media (n=5)||101-113||107|
Samples were spiked with high concentrations of Human CCNB 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 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.