Human Hepatitis B Surface Antigen (HBsAg) ELISA Kit

Product OverviewHepatitis B Surface Antigen (HBsAg) ELISA Kit is an ELISA Kit for the in vitro quantitative measurement of Human Hepatitis B Surface Antigen (HBsAg) concentrations in serum, plasma, tissue homogenates, cell lysates and other biological fluids.Introduction Hepatitis B virus (HBV) is a hepadnavirus that causes damage to the liver in humans. Like other hepadnaviruses, its genome is comprised of circular DNA, with some of it being single-stranded, and the rest double-stranded. The virus enters hepatocytes and replicates within them using a complex series of events, including a reverse-transcription step. Symptoms of infection include malaise, nausea, jaundice, hepatic fibrosis, liver cancer, and acute liver failure. The disease is prevented using a widely-available vaccine, and infections are treated with nucleoside analogues and other antiretrovirals. Target Hepatitis B Surface Antigen (HBsAg) Reactivity Human Tested Applications ELISA Recommended dilutions Optimal dilutions/concentrations should be determined by the end user. Storage Shipped at 4 °C. Upon receipt, store the kit according to the storage instruction in the kit's manual. Validity The validity for this kit is 6 months. Stability The stability of the kit is determined by the rate of activity loss. The loss rate is less than 5% within the expiration date under appropriate storage conditions. To minimize performance fluctuations, operation procedures and lab conditions should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same user throughout. Test Range 0.31 ng/ml - 20 ng/ml Sensitivity 0.19 ng/ml Standard Form Lyophilized Detection Method Colorimetric Assay Type Sandwich Assay Data Quantitative Sample Type Serum, plasma, tissue homogenates, cell lysates and other biological fluids. Assay Principle This kit is based on sandwich enzyme-linked immuno-sorbent assay technology. An antibody is pre-coated onto a 96-well plate. Standards, test samples, and biotin-conjugated reagent are added to the wells and incubated. The HRP-conjugated reagent is then added, and the whole plate is incubated. Unbound conjugates are removed using wash buffer at each stage. TMB substrate is used to quantify the HRP enzymatic reaction. After TMB substrate is added, only wells that contain sufficient HbsAg will produce a blue coloured product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow colour is proportional to the HbsAg amount bound on the plate. The Optical Density (OD) is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of HbsAg can be calculated. Kit Components The kit components listed are for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product. Pre-coated 96-Well Microplate Standard Standard Diluent Buffer Wash Buffer Detection Reagent A Detection Reagent B Diluent A Diluent B TMB Substrate Stop Solution Plate Sealer Material Required But Not Provided 37°C incubator Multi and single channel pipettes and sterile pipette tips Squirt bottle or automated microplate washer 1.5 ml tubes Distilled water Absorbent filter papers 100 ml and 1 liter graduated cylinders Microplate reader (wavelength: 450 nm) ELISA Shaker Reagent Preparation This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product. 1) Standard: Prepare the standard with the recommended volume of Standard Diluent Buffer, to make the standard solution. Then use the Standard Diluent buffer to carry out serial dilutions of the standard solution, as instructed in the Protocol. 2) Wash Buffer: Dilute the concentrated Wash Buffer with distilled water, as instructed in the Protocol. 3) Detection Reagent Preparation: Calculate the total volume of working solution required. Dilute Detection Reagent A and Detection Reagent B with Diluent A and Diluent B, respectively, at 1:100. Assay Procedure This procedure is provided for reference only. The product manual may differ slightly. The product should be used as stated on the product manual included and delivered together with the product. 1) Set standard, test samples and control wells. 2) Aliquot 100 µl of diluted standard into the standard wells. 3) Aliquot 100 µl of Standard Diluent buffer into control (zero) well. 4) Aliquot 100 µl of diluted samples into the sample wells. Incubate for 90 mins at 37 °C. 5) Aliquot 100 µl of Detection Reagent A to each well. Incubate for 1 hr at 37 °C. 6) Wash 3 times. 7) Aliquot 100 µl of Detection Reagent B to each well. Incubate for 30 mins at 37 °C. 8) Wash 5 times. 9) Aliquot 90 µl of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C. 10) Aliquot 50 µl of Stop Solution. 11) Measure the OD at 450 nm. Availability Shipped within 5-12 working days. Note This product is for research use only.The range and sensitivity is subject to change. Please contact us for the latest product information. For accurate results, sample concentrations must be diluted to mid-range of the kit. If you require a specific range, please contact us in advance or write your request in your order comments.Please note that our ELISA and CLIA kits are optimised for detection of native samples, rather than recombinant proteins. We are unable to guarantee detection of recombinant proteins, as they may have different sequences or tertiary structures to the native protein. Plate coated with Antibody