HA Tag Antibody

Western Blot: HA Tag Antibody [NBP2-21581] - Analysis in human HEK293T cells overexpressed with Ly6K-HA. Western blot image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NBP2-21581] - Analysis of HA-recombinant protein transfected HeLa cell which fixed with paraformaldehyde, using antibody at 1:500 dilution.

Flow Cytometry: HA Tag Antibody [NBP2-21581] - HEK293T cells overexpressed with Ly6K-HA stained with HA Epitope Tag Antibody (red) or secondary only (black). Flow cytometry image submitted by a verified customer review.

ELISA: HA Tag Antibody [NBP2-21581] - An ELISA plate is coated with 50 uL of a DDDDK Tag conjugated BSA at concentration of 5 ug/mL. The peptide conjugated carrier protein is coated and detected with anti-DDDDK Tag antibody at concentration ranged from 3.1 to 200 ng/mL.

Western Blot: HA Tag Antibody [NBP2-21581] - Non-transfected (-) and transfected (+) 293T whole cell extracts (30 ug) were separated by 12% SDS-PAGE, and the membrane was blotted with HA Tag antibody (NBP2-21581) diluted at 1:3000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

• Reactivity: ET
• Applications: WB, ELISA, Flow, ICC/IF, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

HA Tag Antibody Summary

• Immunogen: The immunogen used to generate this antibody corresponds to HA Tag
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:1000 - 1:10000
  • Flow Cytometry
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:1000
  • Immunoprecipitation 1:100 - 1:500
  • Western Blot 1:1000 - 1:10000
• Application Notes: This HA Tag antibody is validated for Flow cytometry and WB from verified customer reviews.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS (pH 7), 20% Glycerol, 1% BSA
• Preservative: 0.025% Proclin 300
• Purity: Antigen Affinity-purified

Alternate Names for HA Tag Antibody

• HA Epitope Tag
• HA Tag
• HA
• Hemagglutinin

Background

Human influenza hemagglutinin (HA) is a glycoprotein required for the infectivity of the human virus and is expressed as a homotrimer on the surface of the viral capsid (1). As a major antigen of the influenza virus, HA constantly evolves to escape herd immunity via antigenic shift which can lead to pandemics (2). The HA tag (YPYDVPDYA-tag) itself has a molecular weight of 1.1 kDa and is a linear epitope derived from amino acids 98-106 of the HA protein. It is used extensively as a general epitope tag in expression vectors (2, 3). The HA tag is frequently added to the N- or C- terminus of a protein of interest to facilitate protein purification, detection, and labeling using anti-HA antibodies (3, 4). HA Tag is cleaved by caspase 3/7 resulting in total loss of immunoreactivity, making it unsuitable for the study of apoptosis (5).

References

1. Wilks, S., Graaf, M. D., Smith, D. J., & Burke, D. F. (2012). A review of influenza haemagglutinin receptor binding as it relates to pandemic properties. Vaccine, 30(29), 4369-4376. doi:10.1016/j.vaccine.2012.02.076

2. Wu, N. C., & Wilson, I. A. (2019). Influenza hemagglutinin structures and antibody recognition. Cold Spring Harbor Perspectives in Medicine, 10(8). doi:10.1101/cshperspect.a038778

3. Zhao, X., Li, G., & Liang, S. (2013). Several affinity tags commonly used in chromatographic purification. Journal of Analytical Methods in Chemistry, 2013, 1-8. doi:10.1155/2013/581093

4. Kimple, M. E., Brill, A. L., & Pasker, R. L. (2013). Overview of affinity tags for protein purification. Current Protocols in Protein Science, 73(1). doi:10.1002/0471140864.ps0909s73

5. Schembri, L., Dalibart, R., Tomasello, F., Legembre, P., Ichas, F., & Giorgi, F. D. (2007). The HA tag is cleaved and loses immunoreactivity during apoptosis. Nature Methods, 4(2), 107-108. doi:10.1038/nmeth0207-107

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Reviews for HA Tag Antibody (NBP2-21581) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Human.

reviewed by: Verified Customer Flow Human 07/06/2016

Summary

• Application: Flow Cytometry
• Sample Tested: human HEK293T cells overexpressed with Ly6K-HA
• Species: Human
• Lot: 41423

reviewed by: Verified Customer WB Human 06/17/2016

Summary

• Application: Western Blot
• Sample Tested: human HEK293T cells overexoressed with Ly6K-HA
• Species: Human
• Lot: 41423

FAQs for HA Tag Antibody (NBP2-21581). (Showing 1 - 1 of 1 FAQ).

1. We are looking for a NON-RABBIT polyclonal anti-HA (hemagglutinin - YPYDVPDYA ) antibody for immunohistochemistry that works in 4% paraformaldehyde-fixed tissue. We have a transgenic mouse expressing an HA-tagged protein and want to perform double-labeling with another antibody made in rabbit. We have had no luck using mouse or rat monoclonal anti-HA antibodies. The non-rabbit polyclonal antibody would preferably be made in goat, although if you have other species let us know. It could be unconjugated or conjugated (eg fluorochrome or HRP). However, we are only interested in a product with demonstrated effectiveness for IHC in a published paper. Do you have any such products?
   • We do have other host species for this target and some that have been mentioned in publications. Unfortunately for our goat antibodies against this tag none of them have been validated in IHC-P, and for two we have shown they do not work. Please see this link to our goat anti-HA Epitope Tag antibodies. Since they work in ICC/IF I would assume as long as the tag is expressed in your tissues that you should still pick it up. In this case we have an Innovators Reward Program you can take advantage of for testing a new application to save some money. Here are the chicken anti-HA Epitope Tag antibodies. None of them have been tested in IHC-P as before, or that would be indicated with a positive result and the application on the datasheet or a negative results with the (-) beside the application. Another option for you as well would be going with a Rabbit polyclonal that we have available that has been well publicized and reviewed by customers (cat# NB600-363). I know you plan on double staining, but one way around this would be to use your other antibody and incubate with the secondary for detection first. After doing that you could have this one directly conjugated to your preferred detection probe and then pick up both of their signals since you would already have bound secondary to your first rabbit primary, there would not be any non-specific binding.