Human HPV16 L1 Ab ELISA Kit

Catalog No.: BEK1257

Size: 96 T

Storage and Expiration: Store at 2-8℃ for 6 months, at -20℃ for 12 months. Store HRP conjugated antibody at 4℃ for one year.

Application: For qualitative detection of HPV16 L1 Ab in Human serum, plasma or cell culture supernatant.

Introduction

Papillomaviridae is an ancient taxonomic family of non-enveloped DNA viruses, collectively known as papillomaviruses. Antibodies to HPV16 proteins are associated with increased risk for HPVOPC. Among patients with OPC, HPV16 Abs are associated with tumor HPV status, in particular among HPV positive patients with no or little smoking history. The amino (N) terminus of the human papillomavirus (HPV) minor capsid protein L2 can induce low-titer, cross-neutralizing antibodies. Immunization with HPV16 L1-HPV16 L2 (chimera 17-36) VLP in adjuvant applicable for human use induces broad-spectrum neutralizing antibodies against HPV types evolutionarily divergent to HPV16 and thus may protect against infection with mucosal high-risk, low-risk, and beta HPV types and associated disease.

Principle of the Assay  

This kit was based on indirect enzyme-linked immune-sorbent assay technology. The recombinant HPV16 L1 antigens were pre-coated onto 96-well plates. The test samples were added to the wells and incubated subsequently, the HPV16 L1 antibodies in the test samples combined to the pre-coated antigens specifically, then followed by washing with wash buffer to remove unspecific components. And the HRP conjugated antibodies were added and incubated to bind to human antibodies in plate, the unbound conjugates were washed away with wash buffer. TMB substrate were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. Read the O.D. absorbance at 450 nm in a microplate reader.

Kit components

1. One 96-well plate pre-coated with Human HPV16 L1 antigen

2. Negative control: 0.5 ml

3. Positive control: 0.5 ml

4. Sample diluent buffer: 30 ml

5. HRP conjugated anti-Human antibody (Concentrated): 130 μl. Dilution: 1:100 (Store at 4℃, do NOT store it at -20℃).

6. Antibody diluent buffer: 12 ml

7. TMB substrate: 10 ml

8. Stop solution: 10 ml

9. Wash buffer: 30 ml (25x). Dilution: 1:25

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Absorbent filter papers
8. Plastic or glass container with volume of above 1L