Influenza A H3N2 Lysate | H3N2 lysate

Influenza A H3N2 (A/Kansas/14/2017) Neuraminidase/NA Insect Cell Lysate (WB positive control)

• Applications: Western Blot
• Synonyms: Influenza A H3N2; Influenza A H3N2 (A/Kansas/14/2017) Neuraminidase/NA Insect Cell Lysate (WB positive control); H3N2 NA Overexpression Lysate; H3N2 lysate

• Host: Baculovirus-Insect Cells
• Form/Format: Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.; 1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).

• Applicable Applications for H3N2 lysate: Western Blot (WB)
• Species: H3N2
• Sequence Information: A DNA sequence encoding the Influenza A virus (A/Kansas/14/2017 (H3N2)) neuraminidase (AVG71505.1)(Ile30-Ile469) was expressed with a vasodilator-stimulated phosphoprotein tetramerization domain at the N-terminus 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 (Sigma). 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 pH6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
• 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.
• 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.

Related Product Information for H3N2 lysate

Description: 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.

Background: 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. We have 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.

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.