Influenza A H5N1 Hemagglutinin/HA Cell Lysate | HA cell lysate
Influenza A H5N1 (A/whooper swan/Mongolia/244/2005) Hemagglutinin/HA HEK293 Cell Lysate (WB positive control)
Applications
Western Blot
Synonyms
Influenza A H5N1 Hemagglutinin/HA; Influenza A H5N1 (A/whooper swan/Mongolia/244/2005) Hemagglutinin/HA HEK293 Cell Lysate (WB positive control); Harvey rat sarcoma viral oncogene homolog; HA cell lysate
Host
HEK293 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
524aa
Sequence
Met1-Gln531
Applicable Applications for HA cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
H5N1
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.
Sequence Construction
A DNA sequence encoding the extracellular domain (Met 1-Gln 531) of the influenza hemagglutinin (A/whooper swan/Mongolia/244/2005 (H5N1)) (ACZ36881.1), (HA1+HA2, uncleaved, cleavage site mutated (RRRRKR-TETR)) was expressed, fused with a C-terminal polyhistidine tag.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells 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 HA cell lysate
The influenza viral Hemagglutinin (HA) protein is a homotrimer with a receptor binding pocket on the globular head of each monomer.HA has at least 18 different antigens. These subtypes are named H1 through H18.HA has two functions. Firstly, it allows the recognition of target vertebrate cells, accomplished through the binding to these cells' sialic acid-containing receptors. Secondly, once bound it facilitates the entry of the viral genome into the target cells by causing the fusion of the host endosomal membrane with the viral membrane. The influenza virus Hemagglutinin (HA) protein is translated in cells as a single protein, HA, or hemagglutinin precursor protein. For viral activation, hemagglutinin precursor protein (HA) must be cleaved by a trypsin-like serine endoprotease at a specific site, normally coded for by a single basic amino acid (usually arginine) between the HA1 and HA2 domains of the protein. After cleavage, the two disulfide-bonded protein domains produce the mature form of the protein subunits as a prerequisite for the conformational change necessary for fusion and hence viral infectivity.
This H5N1 Hemagglutinin/HA overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Hemagglutinin/HA protein from the overexpression lysate was verified.
This H5N1 Hemagglutinin/HA overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Hemagglutinin/HA protein from the overexpression lysate was verified.
References
White JM, Hoffman LR, Arevalo JH, et al. Attachment and entry of influenza virus into host cells. Pivotal roles of hemagglutinin. In Chiu W, Burnett RM, Garcea RL. Structural Biology of Viruses.1997
Suzuki Y.Sialobiology of influenza: molecular mechanism of host range variation of influenza viruses. Biol. Pharm. Bull. 2005.
Senne DA, Panigrahy B, Kawaoka Y, et al. Survey of the hemagglutinin (HA) cleavage site sequence of H5 and H7 avian influenza viruses: amino acid sequence at the HA cleavage site as a marker of pathogenicity potential. Avian Dis. 1996
Donald J. Benton, Influenza hemagglutinin membrane anchor, PNAS, 2018
Suzuki Y.Sialobiology of influenza: molecular mechanism of host range variation of influenza viruses. Biol. Pharm. Bull. 2005.
Senne DA, Panigrahy B, Kawaoka Y, et al. Survey of the hemagglutinin (HA) cleavage site sequence of H5 and H7 avian influenza viruses: amino acid sequence at the HA cleavage site as a marker of pathogenicity potential. Avian Dis. 1996
Donald J. Benton, Influenza hemagglutinin membrane anchor, PNAS, 2018
NCBI and Uniprot Product Information
Molecular Weight
The secreted recombinant influenza A H5N1 HA (A/whooper swan/Mongolia/244/2005 (H5N1)) comprises 524 amino acids and has a predicted molecular mass of 59.7kDa.
As a result of glycosylation, it migrates as an approximately 65-75kDa band in SDS-PAGE unde
As a result of glycosylation, it migrates as an approximately 65-75kDa band in SDS-PAGE unde
Protein Family
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Product Notes
The HA (Catalog #AAA8114712) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 524aa. AAA Biotech's Influenza A H5N1 Hemagglutinin/HA can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the HA for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Gln53 1. It is sometimes possible for the material contained within the vial of "Influenza A H5N1 Hemagglutinin/HA, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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