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Influenza A H5N1 (A/Anhui/1/2005) HA / Hemagglutinin / HA Insect Cell Lysate (WB positive control)

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H5N1 HA Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Baculovirus-Insect cells
Product Description:Baculovirus-Insect Cell lysate that Influenza A H5N1 (A/Anhui/1/2005) HA / Hemagglutinin transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
Sequence information:A DNA sequence encoding the extracellular domain of Influenza A virus (A/Anhui/1/2005(H5N1) (ABD28180.1) hemagglutinin (Met 1-Gln 527,Ala 150Val) with cleavage site mutated (RERRRKR→TETR, HA1+HA2, uncleaved) was expressed with the bacteriophage T4 fibritin and a polyhistidine tag at the C-terminus.
Predicted N Terminal:Asp 17
Molecule Mass:The secreted recombinant hemagglutinin of Influenza A virus (A/Anhui/1/ 2005(H5N1) comprises 557 amino acids with the predicted molecular mass of 63 kDa. As a result of glycosylation, it migrates as an approximately 66 kDa band in SDS-PAGE under reducing conditions.This recombinant HA proteins are polymers based on HPLC analysis (data not shown).
H5N1 HA Transfected / Overexpression Cell Lysate Usage Guide
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 (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis 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.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
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 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
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Catalog: 11048-V08BL-300
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