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?

Name: HA Tag Antibody
Brand: Novus Biologicals
SKU: NBP2-21581
Rating: 5 (2 reviews)

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.

HA Tag Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-21581

(2)

Product Datasheet / COA

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Conjugate

Catalog #

HRP

NBP2-16773

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species Reactivity

Epitope Tag

Applications

Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP)

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

View all HA Tag Antibodies »

Product Specifications

Immunogen

The immunogen used to generate this antibody corresponds to HA Tag

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Novus Biologicals Rabbit HA Tag Antibody (NBP2-21581) is a polyclonal antibody validated for use in WB, ELISA, Flow, ICC/IF and IP. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for HA Tag Antibody

Western Blot: HA Tag Antibody [NBP2-21581]

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]

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]

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.

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Applications for HA Tag Antibody

Application

Recommended Usage

Chromatin Immunoprecipitation (ChIP)

Assay Dependent

ELISA

1:1000 - 1:10000

Flow Cytometry

Validated by a verified customer review

Immunocytochemistry/ Immunofluorescence

1:100 - 1:1000

Immunoprecipitation

1:100 - 1:500

Western Blot

1:1000 - 1:10000

Reviewed Applications

Read 2 reviews rated 5 using NBP2-21581 in the following applications:

Flow Cytometry (1 Review)
Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

PBS, 20% Glycerol

Preservative

0.025% Proclin 300

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: HA Tag

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

Alternate Names

HA Epitope Tag

Additional HA Tag Products

Product Documents for HA Tag Antibody

Product Datasheets

Download Product Datasheets

COA

Product Specific Notices for HA Tag Antibody

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.

Citations
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HA Tag Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-21581

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

Concentration

View all HA Tag Antibodies »

Product Specifications

Immunogen

Clonality

Host

Isotype

Description

Scientific Data Images for HA Tag Antibody

Western Blot: HA Tag Antibody [NBP2-21581]

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody [NBP2-21581]

Flow Cytometry: HA Tag Antibody [NBP2-21581]

ELISA: HA Tag Antibody [NBP2-21581]

Western Blot: HA Tag Antibody [NBP2-21581] -

Applications for HA Tag Antibody

Chromatin Immunoprecipitation (ChIP)

ELISA

Flow Cytometry

Immunocytochemistry/ Immunofluorescence

Immunoprecipitation

Western Blot

Reviewed Applications

Formulation, Preparation, and Storage

Purification

Formulation

Preservative

Concentration

Shipping

Stability & Storage

Background: HA Tag

Alternate Names

Additional HA Tag Products

Product Documents for HA Tag Antibody

Product Datasheets

COA

Certificate of Analysis

Product Specific Notices for HA Tag Antibody