Human RGC32(Response Gene To Complement 32)ELISA Kit (HUES03399)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||0.31-20 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human RGC32 in samples. No significant cross-reactivity or interference between Human RGC32 and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human RGC32. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human RGC32 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 RGC32, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human RGC32. The concentration of Human RGC32 in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||RGC32: modulates the activity of cell cycle-specific kinases. Interacts with CDK1 and PLK1. Enhances CDK1 activity and contribute to the regulation of the cell cycle. May inhibit growth of glioma cells by promoting arrest of mitotic progression at the G2/M transition. Fibrogenic factor contributing to the pathogenesis of renal fibrosis through fibroblast activation. Interacts with SMAD3. Cytoplasmic in unstimulated cells; translocates to the nucleus following activation by complement. Associated with the centrosome during prometaphase and metaphase. Induced by Epstein-Barr virus (EBV). Up-regulated in aorta endothelial cells in response to complement activation. Important in vascular smooth muscle cell (VSMC) differentiation following transforming growth factor-beta (TGF-beta) induction. Its expression is greatly induced by TGF-beta in neural crest cells, leading to the induction of VSMC gene promoters. Smad and RhoA signaling are important for RGC-32 activation in neural crest cells. Forms a complex with hnRNP A/B that regulates the transcription of VSMC differentiation genes. Two isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:Protein kinase, regulatory subunit; Cell cycle regulation
Chromosomal Location of Human Ortholog: 13q14. 11
Cellular Component: centrosome; cytoplasm; nucleus
Molecular Function:protein binding; protein kinase activator activity; protein kinase binding
Biological Process: positive regulation of mitosis; positive regulation of collagen biosynthetic process; negative regulation of cytokine secretion; negative regulation of blood vessel endothelial cell migration; G1/S-specific positive regulation of cyclin-dependent protein kinase activity; complement activation; negative regulation of cell proliferation; negative regulation of angiogenesis; negative regulation of exit from mitosis; positive regulation of stress fiber formation; negative regulation of endothelial cell proliferation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription factor activity; positive regulation of cytokine secretion
|NCBI Summary:||This gene is thought to regulate cell cycle progression. It is induced by p53 in response to DNA damage, or by sublytic levels of complement system proteins that result in activation of the cell cycle. The encoded protein localizes to the cytoplasm during interphase and to centrosomes during mitosis. The protein forms a complex with polo-like kinase 1. The protein also translocates to the nucleus in response to treatment with complement system proteins, and can associate with and increase the kinase activity of cell division cycle 2 protein. In different assays and cell types, overexpression of this protein has been shown to activate or suppress cell cycle progression. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||132626811|
|NCBI Gene ID:||28984|
|NCBI Accession:||NP_054778. 2|
|UniProt Secondary Accession:||Q9H4X1,Q6NZ48, Q9UL69,|
|UniProt Related Accession:||Q9H4X1|
|Molecular Weight:||12,924 Da|
|NCBI Full Name:||regulator of cell cycle RGCC|
|NCBI Synonym Full Names:||regulator of cell cycle|
|NCBI Official Symbol:||RGCC|
|NCBI Official Synonym Symbols:||RGC32; RGC-32; C13orf15; bA157L14. 2|
|NCBI Protein Information:||regulator of cell cycle RGCC|
|UniProt Protein Name:||Regulator of cell cycle RGCC|
|UniProt Synonym Protein Names:||Response gene to complement 32 protein; RGC-32|
|Protein Family:||Regulator of cell cycle|
|UniProt Gene Name:||RGCC|
|UniProt Entry Name:||RGCC_HUMAN|
As the OD 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 RGC32 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human RGC32 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.88||5.49||3.58||7.22||4.66||4.56|
The recovery of Human RGC32 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)||86-100||92|
|Cell culture media (n=5)||83-96||90|
Samples were spiked with high concentrations of Human RGC32 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 ELISA 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||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.