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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||Not Applicable|
|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.
|Immunoprecipitation (IP)||See 1 publications below|
26183-1MG 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.
|Not Applicable||60 µg/mL||
A high-affinity recombinant antibody permits rapid and sensitive direct detection of myeloperoxidase.
26183 was used in immunoprecipitation to evaluate a recombinant myeloperoxidase antibody
|McDonnell B,Hearty S,Finlay WJ,O'Kennedy R||Analytical biochemistry (410:1)||2011|