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Influenza A H5N1 (A/Hubei/2011) Hemagglutinin (HA1 Subunit) HEK293 Cell Lysate (WB positive control)

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H5N1 HA Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Human Cells
Product Description:Human Cell lysate that Influenza A H5N1 (A/Hubei/2011) HA1 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 N-terminal segment (Met 1-Arg 345) of the influenza hemagglutinin (A/Hubei/2011 (H5N1)), termed as HA1, was fused with a polyhistidine tag at the C-terminus.
Predicted N Terminal:Asp 17
Molecule Mass:The secreted recombinant HA1 subunit of influenza A H5N1 HA (A/Hubei/2011 (H5N1)) comprises 340 amino acids and has a predicted molecular mass of 38.8 kDa. As a result of glycosylation, it migrates as an approximately 50 kDa band in SDS-PAGE under reducing conditions.
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.
All information of our products is subject to change without notice. Please refer to COA enclosed in shipped package for the newest information.