Product Informations

• Product No. BDAA0131
• Product Name HPV 16 pseudovirus (GFP)
• Category 651
• Dilution Ratio 参考稀释倍数: 1:500
• Shipping and Storage Conditions 冻存于-75+5*C以下，有效期大于12个月
• Precautions 1.我们提供的 HPV 假病毒为复制缺陷型病毒，即该病毒感染目的细胞后不会利用宿主细胞产生新的病毒颗粒； 2.实验操作需要在生物安全柜条件下进行，并穿戴好实验服、口罩和手套等个人防护用品； 3.如果实验时本品不慎溅出，请立即使用 84 消毒液对其进行灭活处理，如果溅到眼睛、皮肤或其他身体部位请立即使用大量清水冲洗； 4.使用本品所产生的实验废弃物需要通过高压灭菌处理后按照医疗废弃物处理要求进行处理。
• Instruction Manual /files/uploads/instructions/2024/202408/BDAA0129_BDAA0143_HPV假病毒(GFP)_产品使用说明书240823.pdf
• Application 可以用于HPV抗血清或者中和抗体滴度检测
• Key Ingredients Brij58、Benzonase、Plasmid Safe、DPBS-Mg、NaCI、葡萄糖、磷酸二氢钾、磷酸氢二钠、氯化钾和假病毒。

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Product Details

【 Product Introduction】
Human Papillomavirus (HPV for short) is mainly transmitted through close human contact, such as sexual contact. It can cause various proliferative epithelial lesions in humans, including papillomas (warts) and tumor-like lesions. The related fatal malignant tumors and various contagious diseases it causes pose a serious threat to human health. The neutralizing antibody level of HPV is a key indicator for HPV-related research and development.
Boao Long selected relevant genes from the HPV L1 and L2 gene sequences publicly available on Genbank, constructed gene expression plasmids, and then co-transfected them with gene reporter plasmids into 293FT cells to construct HPV pseudoviruses. This virus contains the capsid protein of HPV virus, and the DNA sequence of green fluorescent protein (GFP) is encapsulated within the virus.

【 Intended Use 】
It can be used for the detection of HPV antiserum or neutralizing antibody titers

【 Pseudovirus infecting cells Picture (Example)】

 
 


 【 Main Ingredients】
Brij58, Benzonase, Plasmid Safe, DPBS-Mg, NaCI, glucose, potassium dihydrogen phosphate, disodium hydrogen phosphate, potassium chloride and pseudovirus.

【 Storage Conditions and Validity Period
Store at a temperature below -75±5℃ and have a validity period of more than 12 months.

【 Usage Method 】
Collect 293FT cells in the logarithmic growth phase, count them with a hemocytometer, and dilute the cells to a concentration of 1.5× 105 cells /mL with DMEM complete culture medium (DMEM high glucose medium containing 10% fetal bovine serum, 0.1mM non-essential amino acids, 6mM L-glutamine, 1mM sodium pyruvate, and 1% penicillin streptomycin). Inoculate the cells into 96-well cell culture plates at 100μl per well. Incubate in a CO2 incubator for 4 to 6 hours and then use for pseudovirus infection.
2. Dilution of pseudovirus: Take out the frozen pseudovirus and place it on ice to melt or at 4±1℃ or room temperature to melt naturally. After it is completely melted, dilute the pseudovirus to the working concentration using DMEM complete medium (containing 10% fetal bovine serum). Due to the varying infection efficiency of pseudoviruses on different cells, it is recommended to conduct a pre-experiment before the formal test to determine the optimal viral load (the reference titer is indicated on the product label, and the preferred virus dilution ratio = reference titer /2).
3. Premixing of the sample to be tested and pseudovirus: Take a new 96-well cell culture plate, add 55μl of sample and 55μl of pseudovirus diluent to each well, gently tap around to mix well, and at the same time set up a pseudovirus control well with complete culture medium replacing the sample and a blank control well without sample or pseudovirus.
4. Pre-incubation of the sample to be tested and the pseudovirus mixture: Place the 96-well culture plate in a 4 ° C refrigerator for 1 hour.
5. Pseudovirus infection: Take 100μL of the pseudovirus and the sample composition to be tested, as well as the control, from each well of the 96-well culture plate. Slowly add them to the corresponding Wells of the culture plate that have been pre-lined with cells, and gently pat the edges of the culture plate to mix well.
6. Infection detection: On the third day of cell infection with pseudovirus, the infection efficiency is determined by observing the activity of green fluorescent protein expression.
7. To better maintain the product's activity, the pseudo-virus product should minimize repeated freezing and thawing. It is recommended that after receiving the product, the customer dilute it to 1ml with DMEM complete medium (containing 10% fetal bovine serum), then aliquot it into 50μl small packages and store them at -75±5℃. Use it in several portions (or aliquot according to the customer's own experimental needs).

【 Notes】
1. The HPV pseudovirus we provide is a replication-defective virus, meaning that after infecting the target cells, it will not use the host cells to produce new virus particles.
2. Experimental operations must be carried out under the condition of a biosafety cabinet, and personal protective equipment such as laboratory coats, masks and gloves must be worn properly.
3. If this product accidentally splashes during the experiment, please immediately use 84 disinfectant to inactivate it. If it splashes into the eyes, skin or other parts of the body, please rinse immediately with plenty of water.
The experimental waste generated from the use of this product needs to be sterilized under high pressure and then disposed of in accordance with the requirements for medical waste treatment.
5. The titer definition method for the Boao Long HPV pseudovirus product: Use the instrument reading (enzyme-linked immunofluorescence spot analyzer, CTL, S6 Universal). When the reading is 700 (which can be expressed as 700 cells within the detection range of the instrument having fluorescence), the current dilution factor of the pseudovirus is the titer value of this batch of products.