Anti-Vaccinia virus L1 antibody, mouse monoclonal (NP3), neutralizing
Product code65-039

Size

50 µg

Storage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from hybridoma cell culture medium.

ImmunogenPurified Recombinant Vaccinia virus nucleoprotein (L1) expressed in E. coli.

Isotype

Mouse IgG1κ

Reactivity

Nucleoprotein (L1 protein) of vaccinia virus and cowpox virus

Special notes

neutralizing antibody

Application

1. Western blotting: 1/400-800 (Fig.1)
2. Immunofluorescence: 1/400 (Fig.2)

Background

Variola virus (VAV), vaccinia virus (VV), cowpox virus (CPV) and Monkeypox(MPV) belong to the genus Orthopoxvirus of the family Poxviridae. The VAV andMPV cause serious, contagious, and sometimes fatal disease. Therefore,
confirmation of these outbreaks requires rapid and reliable detection anddiagnosis.
It has been shown that several major antigens are induced in cells infected with
poxviruses, i.e., the nucleoprotein (NP) antigen, neutralization (NT)-associatedantigen. The NP (also called L1 or L1R protein) is a so-called common antigen,as shown by cross-reactivity with cells infected with almost all poxviruses. TheL1 protein, a component of the mature virion membrane, is required for virus
entry into host cells and is a target for neutralizing antibody. The L1 is a 250amino-acid polypeptide (with molecular weight of 28kDa). Although L1 has a Cterminal hydrophobic segment embedded in the viral membrane, the 185residue (22kDa), disulfide-bonded ectodomain is located in the cytoplasm before
lysis of the host cell.

Data Link

UniProKB: Q71TT2＿9POXVPlease note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 65-039 Anti-Vaccinia virus L1 antibody, mouse monoclonal (NP3), neutralizing

Fig.1. SDS-PAGE (a) of the recombinant VV L1 protein and Western blot (b) using NP3 antibody.
The recombinant VV L1 protein (185 residues, 22kDa) and the lysates of virus-infected RK13 cells
were applied to SDS-PAGE. (M) Marker, (R) recombinant L1 protein, (V) VV Lister strain, (C) CPV
LB red strain. The NP3 antibody was used at 1/400 dilution. The HRP-conjugated goat anti-mouse
IgG (Abcam) was used at 1/3,000 as the second antibody.

Fig.2. Immunofluorescence staining of VV- or CPV- infected RK13 cells.

Virus-infected anduninfected RK13 cells on a slide glass were fixed with ethanol. (a) VV Lister strain, (b) VV Ikedastrain, (c) CPV LB red strain and (d) uninfected RK13 cells. The NP3 antibody was used at 1/400dilution.

The FITC-conjugated goat anti-mouse IgG (Abcam) was used at 1/3,000 as the secondantibody.

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BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/NT inhibition (%)

100
80

6040
20011040

160

640 2560

PBSDilution of antibody

Fig.3 Neutralization test (NT) of VV by NP3 antibiody
About 100 PFU/ml of virus was incubated for 60 min at 37℃with fourfold serial dilutions of NP2(●) or rabbit anti-VV mouse serum (■) in MEM (+ 2% calf serum). The RK13 cells were incubatedwith the virus-antibody/sample for 60 min at 37℃. After 48 to 64 hrs of incubation at 37℃, plaques
were visualized by staining with 0.13% crystal violet in 2% ethanol. Inhibition rate (%) was calculated
as compared to complete inhibition (100%) of the antibody.
Reference: This antibody has not yet been used in publication.
Related products
65-038Anti-Vaccinia virus L1 antibody, mouse monoclonal (NP2), (cross-reacts with Monkeypoxvirus)

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Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/