HA Tag Antibody [HRP]

• Reactivity: ET
• Applications: WB, ELISA, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: HRP
• Concentration: LYOPH

HA Tag Antibody [HRP] Summary

• Description: Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
  
  Stabilizer: 10 mg/ml Bovine Serum Albumin (BSA)-Immunoglobulin and Protease free
  Color: Substrate dependent
  Conjugation Chemistry: Periodate
  
  This affinity purified antibody is directed against the HA motif and is useful in determining its presence in various assays
• Immunogen: HA Tag Antibody was purified from whole rabbit serum prepared by repeated immunizations with the epitope tag peptide YPYDVPDYA (114-122) from hemagglutinin influenza conjugated to KLH.
• Specificity: This affinity purified Anti-HA antibody is directed against the HA motif and is useful in determining its presence in various assays. This polyclonal anti-HA tag antibody detects over-expressed proteins containing the HA epitope tag. To date, this antibody has reacted with all HA-tagged proteins tested. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:15000
  • Immunohistochemistry 1:500 - 1:2000
  • Immunohistochemistry-Paraffin 1:10 - 1:500
  • Western Blot 1:1000 - 1:5000
• Application Notes: This product is optimally suited for monitoring the expression of HA-tagged fusion proteins. As such, anti-HA/HA can be used to identify fusion proteins containing the HA epitope. The antibody recognizes the HA epitope tag fused to the amino- or carboxy- termini of targeted proteins, as expressed in many commonly used expression vectors. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and HA containing recombinant proteins. Although not tested, this antibody is likely functional for other immunodetection techniques. Affinity purification of the polyclonal antibody results in very low background levels in assays and low cross-reactivity with other cellular proteins.
  
  NOTE: Do NOT add Sodium Azide (it inhibits HRP irreversibly).
• Readout System:
  • NovaLume Plus Chemiluminescent Reagent LUM-555

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
• Preservative: 0.01% Gentamicin Sulfate
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute with 100 ul deionized water (or equivalent)

Alternate Names for HA Tag Antibody [HRP]

• 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.

FAQs for HA Tag Antibody (NBP1-97358). (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.