Anti-Influenza A Virus Nucleoprotein antibody [AA5H] (ab20343)

Ab20343 positively staining formaldehyde fixed Influenza A infected human 293T cells (red) counterstained with AUF-1 (green). ab20343 was used at 1/1000 with no antigen retrieval.

This image is courtesy of an Abreview submitted by Paul Digard on 24 August 2005. We do not have any further information relating to this image.

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ab20343 staining Influenza A Virus Nucleoprotein in porcine lung tissue by Immunohistochemistry (Formalin-fixed, paraffin-embedded sections).
Host receptor binding assays with H1N1 classical swine strain (A/Sw/Iowa/15/30), a subtype closely related to the human 1918 pandemic influenza virus. Briefly, paraffin embedded 5 µm sections of lung tissues were deparaffinised in xylene and rehydrated by alcohol. Deparaffinised tissue sections were incubated with TPCK trypsin treated swine influenza virus for 24 hours at 37°C. Paradoxically, we found that mammalian H1N1 virus binds more efficiently at 37°C than at the usual 4°C. The sections were washed, blocked with goat serum for 30 minutes, and incubated with ab20343 at a 1/1000 dilution, overnight in a humidified chamber at 4°C. A secondary antibody, FITC-labelled goat anti-mouse IgG was applied at 1/500 dilution for 2 hours at room temperature. After three further washes with TBS, the sections were mounted with ProLong Gold anti-fa

Immunocytochemical immunoflurescence analysis of Formaldehyde-fixed human kidney epithelial cells, labelling Influenza A Virus M1 matrix protein with ab20910 at a dilution of 1/500 incubated fro 1 hour at 18°C in 1% FCS PBS. Blocking was with 1% serum incubated for 30 minutes at 18°C. Secondary was a donkey anti-goat Alexa Fluor® 568 undiluted. Cells were transfected with either GFP-M1 or pCDNA2-NP, as indicated on the left hand side of the figure. 24h later cells were fixed and stained for M1 using ab20910 (red) or NP using ab20343 (grey). The abcam 20910 detected M1 in cells transfected with GFP-M1 but not pCDNA3-NP. Moreover, the level of co-localization between GFP-M1 and ab20910 was quite good.

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ab20343 used at a 1/200 dilution in Flow Cytometry.
Ubiquitination negatively regulates antiviral activity of IFITM3.

A and B, HEK293T cells were transfected overnight with indicated plasmids before a 6-hour infection with influenza virus at a multiplicity of infection of 2.5 and analyzed by flow cytometry. Palm? indicates mutation of Cys-71, Cys-72, and Cys-105 to alanine. Ub? indicates mutation of Lys-24, Lys-83, Lys-88, and Lys-104 to alanine. A, cells expressing IFITM3 constructs were analyzed for the percentage of cells that were infected using ab20343. B, antiviral activity was calculated based on the difference in percentage of infection in HA-IFITM3-positive cells compared with vector control with this value set at 100% antiviral activity. Error bars represent the S.D. of triplicate samples. p values were determined using Student's t test. Data are representative of more than five experiments.