Western blot analysis of cell lysates from CHO and CHO HA stable cells using HA Mouse primary antibody and HRP Goat anti-Mouse secondary antibody (#405306). Direct-Blot™ HRP anti-β-actin (#643807) was used as a loading control.
Western blot analysis of cell lysates from CHO and CHO HA stable cells using HA Mouse primary antibody and HRP Goat anti-Mouse secondary antibody (#405306). Direct-Blot™ HRP anti-β-actin (#643807) was used as a loading control.
Immunofluorescence staining of HeLa cells transfected without (A) or with (B, C) multiple tag plasmid using anti-HA Mouse primary Antibody (Clone 16B12). 2 µg/ml (1:250 dilution) Alexa Fluor® 594 (red) Goat anti-Mouse IgG (Cat. No. 405326) was used as secondary antibody. Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective using KEYENCE BZ-X700 fluorescence microscope. Exposure time (Seconds) for (A) is 1/30, and (B, C) is 1/150. Anti-HA Mouse primary Antibody concentrations for (A, B) is (1/1000 dilution), (C) is (1/2000 dilution).
Immunofluorescence of HA.11 tagged Sbhlp protein. Photo courtesy of J Brown and I Davis, UCSF.
Immunoprecipitation of HA protein from CHO and CHO-HA stable cell extracts using HA Mouse primary antibody. Western blot analysis was performed using Direct-Blot™ HRP anti-HA (#901519). Direct-Blot™ HRP anti-β-actin (#643807) was used as a lysate loading control.
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.
Monoclonal antibody HA.11 was raised against the twelve amino acid peptide CYPYDVPDYASL.
Preparation
Ascites
Concentration
The concentration is not quantified as this product is sold as undiluted crude mouse ascites fluid. The concentration might vary from lot-to-lot and an estimated concentration would be 1-3 mg/mL.
Storage & Handling
Store at -20°C. Upon initial thawing, apportion into working aliquots and store at -20°C. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody.
Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is a dilution of 1:1000 – 1:5000 . For immunocytochemistry, the suggested use of this reagent is a dilution of 1:1000 - 1:2000 is suggested. For immunohistochemistry, the suggested use of this reagent is a dilution of 1:1,000 is suggested. For immunoprecipitation, the suggested use of this reagent is a dilution of 1:150 is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
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
(PubMed link indicates BioLegend citation)
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.
Login / Register 
United States ($) USD
Austria (€) EUR
Belgium (€) EUR
Finland (€) EUR
France (€) EUR
Germany (€) EUR
Ireland (€) EUR
Japan (¥) JPY
Luxembourg (€) EUR
Netherlands (€) EUR
Switzerland (CHF) CHF
United Kingdom (£) GBP
Follow Us