HA Tag Antibody - BSA Free

Western Blot: HA Tag Antibody [NBP1-70006] - Detection of 1.0 ug of 12-Epitope Tag Protein Marker Lysate containing the HA epitope tag. A 4-20% gradient gel was used to resolve the protein by SDS-PAGE. The lysate was transferred to nitrocellulose using standard methods. After blocking, the membrane was probed with anti-HA tag antibody for 1 h at room temperature followed by washes and reaction with a 1:20,000 dilution of IRDye 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 for 30 min at room temperature.

Immunohistochemistry-Paraffin: HA Tag Antibody [NBP1-70006] - Arrowheads point to expression of HA tagged proteins in endothelial cells of mouse aorta. Sections of 4 um were prepared from representative paraffin blocks. Sections were then deparaffinized and rehydrated with xylene and alcohol. Citrate buffer antigen retrieval was performed for 30 min in a boiling jar. Anti-HA was diluted in blocking buffer at 1:2,000 and reacted at 4C overnight followed by signal detection using horseradish peroxidase with DAB as the chromogenic substrate. Tissue was counterstained with Mayer's hematoxylin. Personal Communication, Behzad Yeganeh,U. Manitoba, Winnipeg, Canada.

Affinity Purified anti-HA epitope tag polyclonal antibody detects HA tagged recombinant proteins by IHC on formalin fixed paraffin embedded tissue. Arrowheads point to expression of HA tagged proteins in endothelial cells of mouse aorta. Sections of 4 um were prepared from representative paraffin blocks. Sections were then deparaffinized and rehydrated with xylene and alcohol. Citrate buffer antigen retrieval was performed for 30 min in a boiling jar. Anti-HA was diluted in blocking buffer at 1:2,000 and reacted at 4 C overnight followed by signal detection using horseradish peroxidase with DAB as the chromogenic substrate. Tissue was counterstained with Mayer's hematoxylin. Personal Communication, Behzad Yeganeh,U. Manitoba, Winnipeg, Canada.

Anti-HA epitope tag polyclonal antibody detects HA-tagged recombinant proteins by western blot. Polyclonal Rabbit anti-HA epitope tag, at a 1:2,000 dilution, was used to detect 1.0 ug of 12-Epitope Tag Protein Marker Lysate

• Reactivity: ET
• Applications: WB, ELISA, IHC, IP, ChIP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

HA Tag Antibody - BSA Free Summary

• Description: This affinity purified Anti-HA antibody is directed against the HA motif and is useful in determining its presence in various assays
  
  Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. 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.
• 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:
  • Chromatin Immunoprecipitation (ChIP) 1:10-1:500
  • ELISA 1:10000-1:100000
  • Immunohistochemistry 1:500-1:2000
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation 1:10 - 1:500
  • Western Blot 1:2000-1:10000
• 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, immunohistochemistry, and western blotting against both the immunizing peptide and HA containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation, immunocytochemistry, and 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.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Immunogen affinity purified

Alternate Names for HA Tag Antibody - BSA Free

• 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 (NBP1-70006). (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.