Total cell lysate from CHO (lane 1) and CHO stably transfected with HA tag fused protein (lane 2) (15 µg/lane) were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 1:10,000 purified anti-HA.11 or 1:10,000 Direct-Blot™ HRP anti-HA.11 Antibody. For the purified anti-HA.11 antibody, proteins were visualized using a goat anti-mouse-IgG secondary antibody conjugated to HRP. Direct-Blot™ HRP anti-β-actin Antibody was used as a loading controls (lower). Lane M is the MW ladder.
Total cell lysate from CHO (lane 1) and CHO stably transfected with HA tag fused protein (lane 2) (15 µg/lane) were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 1:10,000 purified anti-HA.11 or 1:10,000 Direct-Blot™ HRP anti-HA.11 Antibody. For the purified anti-HA.11 antibody, proteins were visualized using a goat anti-mouse-IgG secondary antibody conjugated to HRP. Direct-Blot™ HRP anti-β-actin Antibody was used as a loading controls (lower). Lane M is the MW ladder.
The HA tag (hemagglutinin) is an amino acid sequence derived from the human influenza hemagglutinin surface glycoprotein, corresponding to amino acids 98-106. It is commonly used as a tag to facilitate detection, isolation, and purification of proteins. The full amino acid sequence is: YPYDVPDYA.
Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested dilution is 1:1000-1:10000. The optimal dilution should be determined by titration for each individual assay of interest.
25 µl and 100 µl of Direct-Blot™ HRP antibody can be used for approximately 5 and 20 Western blots, respectively, at the recommended concentration/dilution.
Application Notes
Additional tested and reported applications of the 16B12 clone for the relevant formats include: western blot (WB), immunocytochemistry (ICC), immunoprecipitation (IP), and flow cytometry (FC).
*Our Posi-Tag Control Protein (931301) can be used as a helpful positive control for this antibody.
This second-generation HA antibody is an excellent substitute for the 12CA5 monoclonal antibody. The HA.11 antibody recognizes the influenza hemagglutinin epitope (YPYDVPDYA) which has been used extensively as a general epitope tag in expression vectors. The extreme specificity of the antibody allows unambiguous identification and quantitative analysis of the tagged protein. The HA.11 antibody recognizes HA epitopes located in the middle of protein sequences as well as at the N- or C-terminus.
Application References
Kim JY, et al. 2003. J Neurosci. 23:5561. (IP, WB)
Helliwell SB, et al. 2001. J Cell Biol. 153:649. (WB)
This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.
