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  • 產(chǎn)品名稱：HumanCCN3/NOV/IGFBP9ELISAPairSet

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  • 產(chǎn)品廠商：SinoBiological
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HumanCCN3/NOV/IGFBP9ELISAPairSet
詳情介紹：
Purpose The ELISA Pair Set is a solid phase sandwich ELISA (Enzyme-Linked Immunosorbent Assay). It utlizes monoclonal antibody specific for CCN3 / NOV / IGFBP9 coated on a 96-well plate. Standards and samples are addedto the wells, and any CCN3 / NOV / IGFBP9 present binds to the immobilized antibody. The wells are washed and ahorseradish peroxidase conjugated mouse anti-CCN3 / NOV / IGFBP9 monoclonal antibody is then added, producingan antibody-antigen-antibody "sandwich". The wells are again washed and TMB substrate solution is loaded, whichproduces color in proportion to the amount of CCN3 / NOV / IGFBP9 present in the sample. To end the enzymereaction, the stop solution is added and absorbances of the microwell are read at 450 nm.
Sensitivity The minimum detectable dose of Human CCN3 / NOV / IGFBP9 was determined to be approximately 31.25 pg/mL. This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.
Characteristics Pair Set
Components Capture Antibody - 0.5 mg/mL of rabbit anti-NOV monoclonal antibody, Dilute to a workingconcentration of 2.0 μg/mL in CBS before coating.
Detection Antibody - 0.5 mg/mL mouse anti-NOV monoclonal antibody conjugated tohorseradish-peroxidase (HRP) . Dilute to working concentration of 0.05 μg/mL in detection antibodydilution buffer before use.
Standard - Each vial contains 100 ng of recombinant NOV. Reconstitute standard powder with1mL detection antibody dilution buffer. After reconstitution, store at -20 °C to -70 °C in a manualdefrost freezer. A seven-point standard curve usi ng 2-fold serial dilutions in sample dilution buffer,and a high standard of 2000 pg/mL is recommended.
Material not included CBS - 0.05 M
Na2CO3 - 0.05 M
NaHCO3, pH 9.6, 0.2 μm filtered
TBS - 25 mM
Tris, adjust pH to 7.4 by HCl
Wash Buffer - 0.05 % Tween20 in TBS, pH 7.2 - 7.4
Blocking Buffer - 5 % BSA in Wash Buffer
Sample dilution buffer - 0.5 % BSA in wash buffer, pH 7.2 - 7.4, 0.2 μm filtered
Detection antibody dilution buffer - 0.5 % BSA in wash buffer, pH 7.2 - 7.4, 0.2 μm filtered
Substrate Solution: To achieve best assay results, fresh substrate solution is recommended Substrate stock solution - 10 mg/mL TMB (Tetramethylbenzidine ) in DMSO Substrate dilution buffer - 0.05 M Na2HPO4 and 0.025 M citric acid , adjust pH to 5.5
Substrate working solution - For each plate dilute 250 μL substrate stock solution in 25 mL substrate dilutionbuffer and then add 80 μL 0.75 % H2O2 , mix it well
Stop Solution - 2 N H2SO4
Alternative Name NOV
Background Protein NOV homolog, also known as Nephroblastoma-overexpressed gene protein homolog, NOV, and CCN3, issecreted protein which belongs to the CCN family. CCN3 expression was observed in a broad variety of tissues fromthe early stage of development. It contains one CTCK (C-terminal cystine knot-like) domain, one IGFBP N-terminaldomain, one TSP type-1 domain, and one VWFC domain. CCN3, a founding member of the CCN family of growthregulators, was linked with hematology in 2003 when it was detected in human serum. CCN3 acts through the corestem cell signalling pathways including Notch and Bone Morphogenic Protein, connecting CCN3 with the modulation ofself-renewal and maturation of a number of cell lineages including hematopoietic, osteogenic and chondrogenic. AlteredCCN3 expression is associated with numerous solid tumors including glioblastoma, melanoma, adrenocortical tumours, prostate cancer and bone malignancies including osteosarcoma. CCN3 may affect the extracellular environment of theniche for hematopoietic stem cells. CCN3 has emerged as a key player in stem cell regulation, hematopoiesis and acrucial component within the bone marrow microenvironment.
Application Notes Optimal working dilution should be determined by the investigator.
Comment

The Human CCN3 / NOV / IGFBP9 ELISA Pair Set is for the quantitative determination of Human CCN3 / NOV /IGFBP9.This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.
Reagent Preparation
  1. Dilute the capture antibody to the working concentration in CBS. Immediately coat a 96-well microplate with 100 μL per well of the diluted capture antibody. Seal the plate and incubate overnight at 4 °C.
    2. Aspirate each well and wash with at least 300 μL wash buffer, repeating the process two times for a total of three washes. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining wash buffer by inverting the plate and blotting it against clean paper towels.
    3. Block plates by adding 300 μL of blocking buffer to each well. Incubate at room temperature for a minimum of 1 hour.
    4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
  1. Add 100 μL of sample or standards in sample dilution buffer per well. Seal the plate and incubate 2 hours at roomtemperature.
    2. Repeat the aspiration/wash as in step 2 of plate preparation.
    3. Add 100 μL of the detection antibody, diluted in antibody dilution buffer, to each well. Seal the plate and incubate 1hour at room temperature.
    4. Repeat the aspiration/wash as in step 2 of plate preparation.
    5. Add 200 μL of substrate solution to each well. Incubate for 20?minutes at room temperature (if substrate solutionis not as requested, the incubation time should be optimized). Avoid placing the plate in direct light.
    6. Add 50 μL of stop solution to each well. Gently tap the plate to ensure thorough mixing.
    7. Determine the optical density of each well immediately, using a microplate reader set to 450 nm.
Calculation of Results

Calculate the mean absorbance for each set of duplicate standards, controls and samples. Subtract the mean zero standard absorbance from each. Construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. To determine the concentration of the unknowns, find the unknowns' mean absorbance value on the y-axis and draw a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the x-axis and read the concentration. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor. Alternatively, computer-based curve-fitting statistical software may also be employed to calculate the concentration of the sample.
Restrictions For Research Use only
Format Lyophilized
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C/-80 °C
Storage Comment Capture Antibody: Aliquot and store at -20°C to -80°C for up to 6 months from date of receipt. Detection Antibody: Protect it from prolonged exposure to light. Aliquot and store at -20°C to -80°C and for up to 6 months from date of receipt. Standard: Store lyophilized standard at -20°C to -80°C for up to 6 months from date of receipt. Aliquot and store the reconstituted Standard at -80°C for up to 1 month.
Expiry Date 6 months
Supplier Images
image for Human CCN3 / NOV / IGFBP9 ELISA Pair Set (ABIN2010223) Human CCN3 / NOV / IGFBP9 ELISA Pair Set