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Western blot analysis of HA Epitope Tag was performed by various amounts of E. coli lysate containing a multi-epitope tagged protein per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an HA Epitope Tag monoclonal antibody (Product # 26183) at a dilution of 1:10,000 for 1 hour at room temperature on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
|Tested species reactivity||Tag|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||HA peptide YPYDVPDYA derivitized to ovalbumin.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||0.1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
26183 is specific for the HA peptide YPYDVPDYA. The antibody has been used successfully in Western blot and immunoprecipitation applications.
HA Synthetic Peptide (Product # 26184) is available for use in neutralization and control experiments and to competitively elute HA-tagged fusion proteins from immobilized anti-HA affinity resin.
Epitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. An HA tag is frequently incorporated into recombinant proteins for a variety of purposes. An anti-HA antibody can then be used to detect the protein when doing studies with transfected cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
SMYD3 links lysine methylation of MAP3K2 to Ras-driven cancer.
26183 was used in western blot to study the role of MAP3K2 kinase methylation and activation by the SMYD3 lysine methyltransferase in ongogenic Ras signaling
|Mazur PK,Reynoird N,Khatri P,Jansen PW,Wilkinson AW,Liu S,Barbash O,Van Aller GS,Huddleston M,Dhanak D,Tummino PJ,Kruger RG,Garcia BA,Butte AJ,Vermeulen M,Sage J,Gozani O||Nature (510:283)||2014|
Regulation of the transcriptional activation of the androgen receptor by the UXT-binding protein VHL.
26183 was used in immunoprecipitation and western blot to study the mechanism by which the UXT-binding protein VHL modulates androgen receptor transcriptional activation
|Chen S,Chen K,Zhang Q,Cheng H,Zhou R||The Biochemical journal (456:55)||2013|
HA; HA Epitope Tag; HA-tag; human influenza hemagglutinin