Flavivirus E protein antibody

Catalog No. ABIN3200993

Product Details

Target: Flavivirus E protein

Reactivity: Human, Mouse

Host: Mouse

Clonality: Monoclonal

Application: Immunofluorescence (IF), Western Blotting (WB)

Specificity: E protein of Flavivirus

Characteristics: Mouse monclonal generated against flavivirus envelope proteins.  Recognizes Dengue virus, West Nile Virus, Japanese Encephalitis and  Zika virus. It binds to domain II of protein E (fusion loop).  

Purification: Purified from hybridoma supernatant via Protein G

Immunogen: Dengue Virus

Clone: 4G2

Isotype: IgG2a

Application Details

Application Notes: 0.1-1.0 µg/mL

Restrictions: For Research Use only

Independent Validation (ICC)

Validation #101139 (Immunocytochemistry)

by: Division of Clinical Pharmacology, LMU, Munich

No.: #101139

Date: 11/07/2017

Antigen: Flavivirus E protein

Method validated: Immunocytochemistry

Positive Control: 1205Lu human melanoma cells infected with yellow fever vaccine strain YF-17D

Negative Control: Uninfected 1205Lu cells; infected cells without primary antibody

Notes: ABIN3200993 specifically labels the targeted antigen in 1205Lu melanoma cells in ICC. No signal was detected in uninfected negative control tissue and the secondary antibody only control.

Validation Images

1205Lu melanoma cells infected (B, C, D) and uninfected (A) with yellow fever vaccine strain YF-17D were stained with anti-Flavivirus E protein antibody ABIN3200993 and an ER-specific antibody (A, C, D) and AF647- (red) and AF488-conjugated (green) secondary antibodies or only with the secondary antibodies (B). DAPI counterstain (white) was used to reveal cells.

Full Methods

Primary Antibody: ABIN3200993

Secondary Antibody: goat-anti mouse AF647 conjugated antibody (Invitrogen, A21235, lot 1764240)

Full Protocol:

• Grow cells in DMEM supplemented with 10% FCS, 1% L-Glutamin, 1% Pen/Strep (Gibco) to 70% confluency on coverslips coated with Poly L-Lysin (Sigma, P8920).
• Infect cells with yellow fever vaccine strain YF-17D at 2MOI.
• Grow cells on coverslips for 48h.
• Fix cells on coverslips in 4% PFA for 30min at RT.
• Wash cells 2x with PBS.
• Permeabilize cells in PBScontaining 0.1% Triton-X 100 (PBST) for 1h at RT.
• Wash cells 2x with PBS.
• Block non-specific binding with in PBST containing 10% normal goat serum for 2h at RT.
• Incubate slides with primary mouse anti-Flavivirus E protein antibody (antibodies-online, ABIN3200993) diluted 1:200 and rabbit-anti ER antibody (abcam, ab176333, GR265198-3) diluted 1:200 in PBST with 10% NGS ON at 4°C.
• Include wells without primary antibody as negative controls.
• Rinse slides 3x with PBST.
• Incubate slides with secondary goat-anti mouse AF647 conjugatedantibody (Invitrogen, A21235, lot 1764240) diluted 1:1000 and goat-anti rabbit AF488 conjugated antibody (Invitrogen, A-11008, lot 84B2-1) diluted 1:1000 in PBST with 3% NGS for 1h at RT.
• Rinse slide 3x with PBST.
• Nuclear counterstain with DAPI diluted in PBS (Sigma, 10236276001).
• Wash cells 2x with PBS.
• Mount coverslips on glass slides in Mowiol antifade reagent (Calbiochem, 475904).
• Image acquisition with Leica SP5 II confocal imaging system, 20x magnification, 1024x1024 resolution.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: PBS pH 7.4

Storage: -20 °C

Storage Comment: Heat stable , shipped at ambient  temp  Upon delivery aliquot and store in fridge , longterm storage at  -20°C.

References

Tang, Hammack, Ogden, Wen, Qian, Li, Yao, Shin, Zhang, Lee, Christian, Didier, Jin, Song, Ming: "Zika Virus Infects Human Cortical Neural Progenitors and Attenuates Their Growth." in: Cell stem cell, Vol. 18, Issue 5, pp. 587-90, (2017) (PubMed).

Target Details

Target: Flavivirus E protein