H7(Pan) Antigens Hemagglutinin ELISA Development Kit-蚂蚁淘生物

货号： IT-E3Ag-H7-Pan
检测方法： ELISA
数量：中量
供应商： Immune Technology Corp.
规格： 1 kit
H7(Pan) Antigens Hemagglutinin ELISA Development KitCatalog Number: IT-E3Ag-H7-Pan Description: H7(Pan) antigens Hemagglutinin (HA)ELISA Development Kit contains the key componentsrequired for the quantitative analysis of H7(Pan) antigenconcentrations in cell culture supernatants, serum, andother biological samples within the range of0.001-1ng/ml in a sandwich ELISA format. Thecomponents supplied in this kit are sufficient to performthe assay in five 96-well ELISA plates.REAGENTS PROVIDEDCapture Antibody: 100μl of 1mg/ml H7(Pan) anti-HAmonoclonal antibody.HA1(H7N9)(A/Shanghai/2/2013) Standard: 50μl of50μg/ml recombinant HA1(H7N9)(A/Shanghai/2/2013).Detection Antibody: 50μl of biotinylated monoclonalantibody against H7(Pan) HA.Streptavidin-HRP Conjugate: 50μl of HRP-conjugatedstreptavidin.RECOMMENDED MATERIALS & SOLUTIONS*ELISA 96-well plates (Corning Prod # 3590 orequivalent plate)Block Buffer: 5% milk in PBSWash Buffer: 0.05% Tween-20 in PBSDiluent: 0.05% Tween-20, 0.5% milk in PBSSubstrate: TMB Peroxidase SubstrateStop Solution: 2N Sulfuric Acid*Alternatively, these could be purchased under Cat.# IT-200-002— ELISA Plate/Buffer/Substrate Kit.PLATE PREPARATION1. For each 96-well plate, dilute 20μl of CaptureAntibody with 10.5ml of 1xPBS to prepare a coatingsolution. Immediately add 100μl of the coatingsolution to each well. Seal the plate and incubateovernight at 4oC.2. Remove the coating solution by aspirating ordecanting. Invert the plate and blot it briefly againstclean paper towels.3. Add 300μl of Block Buffer to each well. Incubatefor 2 hour at 37oC.4. Aspirate to remove Block Buffer and wash the plate4 times with 300μl of wash buffer per well.ASSAY PROCEDURE1. Standard/Sample: Dilute standard with Diluent toeight concentrations (1ng/ml, 0.316ng/ml, 0.1ng/ml,0.0316ng/ml, 0.01ng/ml, 0.00316ng/ml, 0.001ng/ml,and 0ng/ml). Immediately add 100μl of Standardand sample to each well in triplicate. Incubate at37oC for 1 hour.2. Detection: Aspirate and wash plate 4 times. Dilute10μl of Detection Antibody with 10.5ml of Diluentto prepare a detection solution. Add 100μl of thedetection solution into each well. Incubate at 37oCfor 1 hour.3. Streptavidin Peroxidase: Aspirate and wash plate 4times. Dilute 10μl of Streptavidin-HRP conjugatewith 10.5ml of Diluent. Add 100μl into each well.Incubate at room temperature for 30 minutes.4. Substrate/Stop: Aspirate and wash plate 4 times.Add 100μl of TMB Peroxidase Substrate into eachwell. Incubate at 37oC for 30 minutes. Then add100μl of Stop Solution to each well.5. Read: Determine the optical density of each wellwithin 30 minutes using a microplate reader set to450nm.6. Analysis: Average the triplicate reading for eachstandard, control, and sample, then subtract theaverage zero standard optical density. Create astandard curve by reducing the data using computersoftware capable of generating a four parameterlogistic (4-PL) or other curve-fit. TheHA1(H7N9)(A/Shanghai/2/2013) concentration insample can be determined by regression analysis. Ifsamples have been diluted, the concentrationcalculated from the standard.

货号：	IT-E3Ag-H7-Pan
检测方法：	ELISA
数量：	中量
供应商：	Immune Technology Corp.
规格：	1 kit

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