Influenza A H3N2 Neuraminidase/NA Cell Lysate | NA cell lysate
Influenza A H3N2 (A/Hong Kong/4801/2014) Neuraminidase/NA (His Tag) Insect Cell Lysate (WB positive control)
Applications
Western Blot
Synonyms
Influenza A H3N2 Neuraminidase/NA; Influenza A H3N2 (A/Hong Kong/4801/2014) Neuraminidase/NA (His Tag) Insect Cell Lysate (WB positive control); H3N2 NA Overexpression Lysate; NA cell lysate
Host
Baculovirus-Insect Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
480aa
Sequence
His36-Ile469
Applicable Applications for NA cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
H3N2
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
Sequence Construction
A DNA sequence encoding the Influenza A virus (A/Hong Kong/4801/2014 (H3N2)) neuraminidase (translated amino acids of EPI675797) (His36-Ile469) was expressed with a vasodilator-stimulated phosphoprotein tetramerization domain at the N-terminusa and a polyhistidine tag at the C-terminus.
Preparation Method
Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for NA cell lysate
Neuraminidases are enzymes that cleave sialic acid groups from glycoproteins. Influenza neuraminidase is a type of neuraminidase found on the surface of influenza viruses that enables the virus to be released from the host cell. Influenza neuraminidase is composed of four identical subunits arranged in a square. It is normally attached to the virus surface through a long protein stalk. The active sites are in a deep depression on the upper surface. They bind to polysaccharide chains and clip off the sugars at the end. The surface of neuraminidase is decorated with several polysaccharide chains that are similar to the polysaccharide chains that decorate our cell surface proteins. Neuraminidase (NA) and hemagglutinin (HA) are major membrane glycoproteins found on the surface of the influenza virus. Hemagglutinin binds to the sialic acid-containing receptors on the surface of host cells during initial infection and at the end of an infectious cycle. Neuraminidase, on the other hand, cleaves the HA-sialic acid bondage from the newly formed virions and the host cell receptors during budding. Neuraminidase thus is described as a receptor-destroying enzyme that facilitates virus release and efficient spread of the progeny virus from cell to cell. Influenza antibody and influenza antibodies are very important research tools for influenza identification, influenza vaccine development, and anti-influenza virus therapy development. The monoclonal or polyclonal antibody can be raised with protein based antigen or peptide-based antigen. Antibodies raised with protein-based antigen could have better specificity and/or binding affinity than antibodies raised with peptide based antigen, but the cost associated with the recombinant protein antigen is usually higher. Anti-influenza virus hemagglutinin (HA) monoclonal antibody or polyclonal antibody can be used for ELISA assay, western blotting detection, Immunohistochemistry (IHC), flow cytometry, neutralization assay, hemagglutinin inhibition assay, and early identification of influenza viral infection. MyBioSource has developed state-of-the-art monoclonal antibody development technology platforms: mouse monoclonal antibody and rabbit monoclonal antibody. Our rabbit monoclonal antibody platform is one of a kind and offers some unique advantages over mouse monoclonal antibodies, such as high affinity, low cross-reactivity with rabbit polyclonal antibodies.
This H3N2 Neuraminidase/NA overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of Neuraminidase/NA protein from the overexpression lysate was verified.
This H3N2 Neuraminidase/NA overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of Neuraminidase/NA protein from the overexpression lysate was verified.
References
Sardet C., et al., (1989), Molecular cloning, primary structure, and expression of the human growth factor-activatable Na+/H+ antiporter. Cell 56:271-280.Sardet C., et al., (1990), Growth factors induce phosphorylation of the Na+/H+ antiporter, glycoprotein of 110 kD.Science 247:723-726.Tse C.-M., et al., (1991), Molecular cloning and expression of a cDNA encoding the rabbit ileal villus cell basolateral membrane Na+/H+ exchanger.EMBO J. 10:1957-1967.
NCBI and Uniprot Product Information
Molecular Weight
The recombinant Influenza A virus (A/Hong Kong/4801/2014 (H3N2)) NA consists of 480 amino acids and predicts a molecular mass of 53.9kDa.
Protein Family
