Influenza A Nucleoprotein Antibody | AA5H

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Influenza A Nucleoprotein Antibody | AA5H gallery image 1

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Mouse anti Influenza A Nucleoprotein antibody, clone AA5H used for the detection of viral nucleoproitein in influenza infected HEK-293 cells by western blotting.
Image caption:
Forced expression of influenza viral proteins does not result in impaired IFNβ-induced STAT1 and STAT2 phosphorylation. HEK293 cells were transfected with 500 ng plasmid DNA for expression of viral NP, M, NS, (A) PA, PB1 and PB2 (C) genes (see Table 1 for accession numbers of viral genes) using L2000 according to manufacturer's instructions. Note that the Pol II constructs in use also give rise to expression of second reading frames in the NS, M and PB1 genes (NS2, M2, PB1-F2). 48 h post transfection cells were stimulated with human IFNβ (500 U/ml) for 15 minutes. Total protein lysates were subjected to Western blot analysis using anti-phospho-STAT1, anti-phospho-STAT2, anti-STAT1 antibodies. Expression of influenza viral proteins was monitored with antibodies against NP, M1, NS1, PA, PB1 or PB2. (E) HEK293 cells were infected with the human influenza virus PR8 (H1N1) (MOI = 5) for the indicated time points and were subsequently stimulated for 15 min with either human IFNβ at a concentration of 100 U/ml. Cell lysates were subjected to Western blots as described. (B, D, F) Quantification of relative pSTAT1 and pSTAT2 band intensities in A, C and E using AIDA software and 2D densitometry (Fuji).

From: Pauli E-K, Schmolke M, Wolff T, Viemann D, Roth J, Bode JG, et al. (2008) Influenza A Virus Inhibits Type I IFN Signaling via NF-κB-Dependent Induction of SOCS-3 Expression.
PLoS Pathog 4(11): e1000196.

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Influenza A Nucleoprotein Antibody | AA5H gallery image 2

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Mouse anti Influenza A Nucleoprotein antibody, clone AA5H used for the detection of viral protein in a range of organs from an influenza infected patient using immunohistochemistry on formalin fixed, paraffin embedded tissue sections.
Image caption:
The results of IHC stain with antibody against NP. Positive stain seen with diaminobenzidine (brown; Dako). Slides counterstained with haematoxylin. (A) Positive stain presented in pneumocytes of lung (arrow). (B) Endotheliocyte of aortopulmonary vessel (arrow). (C) Positive stain in nuclei and cytoplasm of axillary lymphoid-node (arrow). (D) Positive stain in lymphocytes of spleen (arrow). (E) Positive stain in cytoplasm of neuron and gliocyte from cerebral cortex (arrows). (F) Positive stain in nuclei and cytoplasm of intestine mucosa (arrows). (G) Positive stain in nuclei of mononuclear-like cells (arrows) with morphological features of macrophages from liver. (H) Positive stain in cytoplasm and nuclei of endotheliocytes of renal contex. (I) Positive stain in nuclei and cytoplasm of epithelial cells from ureter (arrow). Original magnifications: (A) ×20, (B–E) ×40, (F–G) ×20, (H–I) ×40.

From: Gao R, Dong L, Dong J, Wen L, Zhang Y, Yu H, et al. (2010) A Systematic Molecular Pathology Study of a Laboratory Confirmed H5N1 Human Case.
PLoS ONE 5(10): e13315.

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Influenza A Nucleoprotein Antibody | AA5H gallery image 3

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Mouse anti Influenza A Nucleoprotein antibody, clone AA5H used for the identification of influenza viral nucleoprotein in infected cell lysates.
Image caption:
Replication and IFN-β induction by human and avian H5N1 strains in human cells. (A, B) A549 cells were infected at a multiplicity of 0.01 (A) or 1 (B) with the highly-pathogenic human H5N1 IAV isolates A/Hong Kong/156/1997 (HK/97) and A/Thailand/1 (KAN-1)/2004 (Thai/04), the avian H5N1 strains A/chicken/Indonesia/R132/2004 (Ch/Ind), A/duck/Vietnam/TG24-O1/2005 (Duck/VN) and A/common buzzard/Berlin/1/2006 (Buzz/Bln) as well as with the prototypic seasonal A/Panama/2007/99 (Pan/99) H3N2 virus and its mutant variant with a deleted NS1 gene (Pan-delNS1; shown only in panel A). Supernatant of infected cell cultures was sampled at the indicated time points and viral titers were determined by plaque assay. Mean data+SEM of at least three independent experiments is shown. (C) Immunoblot analysis of lysates from A549 cells infected at MOI = 1 using a viral NP-specific antibody at the indicated time points post infection (p.i.). Relative intensities of NP bands normalized to actin controls are depicted in comparison to Thai/04, which was arbitrarily set as 100%. (D, E) Concentrations of IFN-β in cell culture supernatants of A549 cells infected with the indicated viruses at low (MOI = 0,01, panel D) or high multiplicity (MOI = 1, panel E) determined via ELISA at 24 and 48 hpi (D) or at 16 hpi (E). Mean data of at least three independent experiments +/– SEM is shown. Symbols indicate significant differences between given results with p<0.05 (*), p<0.01 (**), or p<0.001(***), respectively, as determined by unpaired two-tailed t-tests.

From: Matthaei M, Budt M, Wolff T (2013) Highly Pathogenic H5N1 Influenza A Virus Strains Provoke Heterogeneous IFN-α/β Responses That Distinctively Affect Viral Propagation in Human Cells.
PLoS ONE 8(2): e56659.

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Influenza A Nucleoprotein Antibody | AA5H gallery image 4

Published customer image:
Mouse anti Influenza A Nucleoprotein antibody, clone AA5H used for the identification of influenza viral nucleoprotein in infected cell lysates.
Image caption:
Published customer image:
Mouse anti Influenza A Nucleoprotein antibody, clone AA5H used for the identification of influenza viral nucleoprotein in infected cell lysates.
Image caption:
H5N1 NS1 proteins inhibit the induction of type I IFN in human cells. (A) IFN-β concentrations of supernatants sampled from A549 cell cultures infected with the Pan99 WT and different NS reassortant viruses (H3N2) (MOI = 0.01) at 24 and 48 hrs p.i., (N=2+/–SEM). (B) Human 293T cells were co-transfected with plasmids expressing the H5N1- or Pan/99 (H3N2)-derived NS segments or empty vector, the human IFN-β promoter reporter plasmid p125-Luc and pRL-TK-Luc to control for transfection efficiency. Subsequently, cells were infected for 16 h with Pan-delNS1 to stimulate the IFN-β promoter before luciferase reporter activity was determined in cell lysates. IFN-β promoter activation in infected cells is shown as x-fold stimulation of firefly luciferase activity compared to transfected and mock infected cells. Mean data of three independent experiments is shown +/– SD. (C) Immunblot detection of viral NP after Pan-delNS1 (H3N2) infection confirmed equal stimulation of the transfected cells analyzed in panel B.

From: Matthaei M, Budt M, Wolff T (2013) Highly Pathogenic H5N1 Influenza A Virus Strains Provoke Heterogeneous IFN-α/β Responses That Distinctively Affect Viral Propagation in Human Cells.
PLoS ONE 8(2): e56659.

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  • Mouse anti Influenza A Nucleoprotein
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  • Product Type
    Monoclonal Antibody
  • Clone
    AA5H
  • Isotype
    IgG2a
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA400IF, P, WBdatasheet pdfdatasheet pdf1 mg
    MCA400
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Influenza A Nucleoprotein antibody, clone AA5H recognizes an epitope within Influenza virus A nucleoprotein. Mouse anti Influenza A Nucleoprotein antibody, clone AA5H can be used in influenza A IFA typing in conjunction with MCA401 (clone GA2B).
    • Intended Use
    • Target Species
      Viral
    • Product Form
      Purified IgG - liquid
    • Reconstitution
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant.
    • Preservative Stabilisers
      0.09%Sodium Azide
    • Immunogen
      Influenza A / Puerto Rico / 8 / 34 (H1N1) and A/Bangkok / 1 / 79 (H3N2) viruses.
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 1.0 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Fusion Partners
      Spleen cells from BALB/c mice were fused with cells of the P3 Ag8.653 mouse myeloma cell line.
    • Storage
      Store at +4oC or at -20oC if preferred.
      This product should be stored undiluted.
      Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      Immunofluorescence
      Immunohistology - Paraffin
      Western Blotting

    • Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional Influenza A Nucleoprotein Antibody Formats

    Formats Clone Applications Sizes available
    Influenza A Nucleoprotein Antibody : Purified AA5H IF, P, WB 1 mg
    • Copyright © 2016 Bio-Rad

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