Immunofluorescent analysis of HeLa cells transfected with a construct containing an HA Epitope Tag. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a DyLight 680-conjugated HA Epitope Tag monoclonal antibody (Product # 26183-D680) at a dilution of 1:25 for at least 1 hour at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or ToxInsight Instrument at 20X magnification.
|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 *|
|Immunofluorescence (IF)||1:25 - 1:100|
|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-D680 has been successfully used in immunofluorescence and Western blot applications. DyLight 680 has an excitation/emission of 692/712 nm.
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-D680 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|