Flavivirus E protein 抗体
Quick Overview for Flavivirus E protein 抗体 (ABIN3200993)
抗原
适用
宿主
克隆类型
应用范围
克隆位点
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特异性
- E protein of Flavivirus
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产品特性
- 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).
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纯化方法
- Purified from hybridoma supernatant via Protein G
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免疫原
- Dengue Virus
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亚型
- IgG2a
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应用备注
- 0.1-1.0 µg/mL
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限制
- 仅限研究用
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- by
- Division of Clinical Pharmacology, LMU, Munich
- No.
- #101139
- 日期
- 2017.11.07
- 抗原
- Flavivirus E protein
- Lot Number
- 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.
- 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.
- Experimental Notes
生效 #101139 (Immunocytochemistry)Validation Images1205Lu 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 -
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状态
- Liquid
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浓度
- 1 mg/mL
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缓冲液
- PBS pH 7.4
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储存条件
- -20 °C
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储存方法
- Heat stable , shipped at ambient temp Upon delivery aliquot and store in fridge , longterm storage at -20°C.
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Zika Virus Infects Human Cortical Neural Progenitors and Attenuates Their Growth." in: Cell stem cell, Vol. 18, Issue 5, pp. 587-90, (2017) (PubMed).
: "
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Zika Virus Infects Human Cortical Neural Progenitors and Attenuates Their Growth." in: Cell stem cell, Vol. 18, Issue 5, pp. 587-90, (2017) (PubMed).
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- Flavivirus E protein
抗原
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