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 with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:500 - 1:1000|
* 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 1 publications below|
26183-BTIN has been successfully used for Western blot and IP.
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||Not Cited||
Valproic acid causes proteasomal degradation of DICER and influences miRNA expression.
26183-BTIN was used in western blot to study the proteasomal degradation of DICER and altered miRNA levels induced by valproic acid
|Zhang Z,Convertini P,Shen M,Xu X,Lemoine F,de la Grange P,Andres DA,Stamm S||PloS one (8:null)||2013|