CVPx ᵀᴹ Human HPV 18 Antigen ELISA Quantitation Kit | Creative Diagnostics DEIASL119 product information

product summary

company name :

Creative Diagnostics

product type :

ELISA/assay

product name :

CVPx ᵀᴹ Human HPV 18 Antigen ELISA Quantitation Kit

catalog :

DEIASL119

quantity :

96T

more info or order :

Creative Diagnostics product webpage

product information

Catalog :

DEIASL119

proudctName :

CVPx ᵀᴹ Human HPV 18 Antigen ELISA Quantitation Kit

common :

HPV

Short Name :

CVPx TM Human HPV 18 Antigen ELISA Quantitation Kit

Detection Method :

sELISA

Common name :

HPV

Species :

Human

Application :

Quantitative

Sample :

Serum and Plasma

Size :

96T

Storage :

The HPV 18 Antigen Standard and HRP-Conjugated Antibody Solution should be stored at -20°C. Other components remain stable under the condition of 2-8°C for six months.

Specificity :

There is no cross reaction whis HPV-16 antigens.

Target :

HPV18

Species Reactivity :

Human

Intended Use :

Enzyme immunoassay for quantitative analysis of HPV-18 antigen in samples.

General Description :

HPV is a double-stranded DNA virus with a circular genome that encodes early genes, including E1, E2, E4, E5, E6, and E7, essential for replication, transcription, and transformation, and late genes L1 and L2, encoding viral capsid proteins. The replication cycle of HPV is tightly linked to the differentiation of the infected epithelium. Indeed, viral protein production and virus assembly occur only in the upper differentiated layers of the epithelium. In the basal layer, HPV particles initially interact with the basement membrane mostly through heparan sulfate proteoglycans (HSPGs)—capsid L1 contacts, and subsequently bind to HSPGs present on the basal keratinocyte cell surface. This attachment triggers conformational changes in the L2 capsid protein, resulting in exposure of a consensus cleavage site in the L2 N-terminus, whose proteolysis facilitates further interaction of the viral capsid with secondary receptor(s) present on the keratinocyte membrane. After such binding, HPVs are generally internalized by clathrin-dependent endocytosis, which initially relies on actin-rich cell protrusions, acting as the transport mechanism along the endocytic machinery.

Principle Of The Test :

This product utilizes the Double Antibody Sandwich Method (Sandwich ELISA). Flat-bottom 96-well plates are coated with anti-HPV-18 antibody. After adding the samples, wash is performed to remove unconjugates. Subsequently, an HRP-labeled anti-HPV18 antibody is added, followed by incubation and wash. Ultimately, the HPV-18 content in the samples can be determined by the extent of TMB color development.

Reagents And Materials Provided :

1. Coated plate: 1 (96 wells) 2. HPV 18 Antigen Standard: Lyophilized, 10 μg/mL, 1 vial. Store at -20°C. 3. HRP-Conjugated Antibody Solution: 100×, 120 μl, 1 vial. Store at -20°C 4. HRP-Conjugated Dilution Buffer: 12 ml, l vial. 5. Sample Diluent: Ready to use, 50 ml, 1 vial. 6. Washing Buffer: 20×, 50 ml, 1 vial. 7. TMB Substrate Solution: 6 ml, 2vial. 8. Stop Solution: 7 ml, 1 vial. 9. Sealing plate film, 2 pieces. 10. Instruction manual

Materials Required But Not Supplied :

1. Microplate reader capable of reading absorbance at 450 nm. 2. Automated plate washer (optional). 3. Pipettes and pipette tips. Multichannel pipettes are recommended for a large numbers of samples. 4. Deionized or distilled water. 5. Test tubes for dilution. 6. ELISA plate shaker 7. 37°C incubator.

Specimen Collection And Preparation :

Use human serum or plasma (citrate, heparin) samples with this assay. If the assay is performed within 5 days after sample collection, the samples should be kept at 2-8°C; otherwise, they should be aliquoted and stored deep-frozen (-60°C). If samples are stored frozen, mix thawed samples well before testing. Avoid repeated freezing and thawing. The serum or plasma samples are recommended to be diluted 10 times, but the actual dilution factor needs to be determined by the customer through pre-testing.

Reagent Preparation :

1. 1× Washing Buffer: Take 1 vial of 20× Washing Buffer, dilute it to 1000 ml with deionized water, mix well for later use. 2. HRP-Conjugated Antibody Working Solution: Dilute the HRP-Conjugated Antibody Solution 100× to prepared HRP-Conjugated Dilution Buffer. Dilution ratio: 1:100, mix well. 3. Standard: Use 200 μL ddH2O solution standard. It is recommended that the standards be prepared no more than 24 hours prior to performing the experiment and should be stored at -20°C. Dilute the Standard with the prepared Sample Diluent. For example: • The remaining standard should be stored at -20°C or below.

Assay Procedure :

1. Equilibrate the required reagents at room temperature (20~25°C) for 30 minutes. 2. Prepare all reagents as previously instructed. 3. Remove excess microplate strips from the plate frame, seal them, and store them vacuum-packed. 4. Add 100 μl of the standard, samples, or control per well. Add 100 μl of the Sample Diluent into the zero well. It is recommended to use at least two replicates of each standard, sample, or control. 5. Cover with the provided plate sealer and incubate for 60 minutes at 37°C in an incubator set at 200~500 rpm. 6. Wash the plate 5 times with the 1× Washing Buffer. 7. Add 100 μL of the prepared HRP-Conjugated Antibody Working Solution to each well. 8. Cover with the provided plate sealer and incubate for 60 minutes at 37°C in an incubator set at 200~500 rpm. 9. Repeat step 6. 10. Add 100 μl of TMB Substrate Solution to each well. Cover with the provided plate sealer and incubate in the dark for 5-10 minutes at room temperature (RT). 11. Add 50 μl of Stop Solution to each well. The color should immediately change to yellow. 12. Within 20 minutes of stopping the reaction, measure the absorbance at 450/630 nm using a microplate reader.

Interpretation Of Results :

Plotting the Standard Curve and Determining the Sample Concentration Using computer reduction software, plot absorbance (linear y-axis) versus concentration (log x-axis) for standards and fit the data with a four-parameter logistic equation. Using the equation of the line, calculate the concentration of analyte in each sample, making sure to correct for any sample dilution.

Detection Range :

7.8～500 ng/ml

Precautions :

1. For professional use only. 2. In case of severe damage to the kit package, please contact CD or your supplier in writing within one week of receiving the kit. Do not use damaged components in test runs, but keep them safe for complaint-related issues. 3. Obey the lot number and expiry date. Do not mix reagents from different lots, and do not use expired reagents. 4. Before starting the assay, read the instructions completely and carefully. Use the valid version of the package insert provided with the kit. Ensure that everything is understood. For further information (clinical background, test performance, automation protocols, alternative applications, literature, etc.), please refer to the local distributor. 5. Follow good laboratory practice and safety guidelines. Wear lab coats, disposable latex gloves, and protective glasses where necessary. 6. Reagents in this kit containing hazardous materials may cause eye and skin irritations. 7. Chemicals and prepared or used reagents must be treated as hazardous waste according to national biohazard safety guidelines or regulations.

applicatons :

Enzyme immunoassay for quantitative analysis of HPV-18 antigen in samples.

Contents of Kit :

Tetramethylbenzidine :