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HA Tag Antibody [HRP]

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-97358

Novus Biologicals, part of Bio-Techne
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NBP1-97358

Key Product Details

Species Reactivity

Epitope Tag

Applications

ELISA, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

HRP

Antibody Source

Polyclonal Rabbit IgG

Concentration

LYOPH mg/ml

Product Specifications

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.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

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

Applications for HA Tag Antibody [HRP]

Application
Recommended Usage

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).

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Reconstitution

Reconstitute with 100 ul deionized water (or equivalent)

Formulation

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 mg/ml

Shipping

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

Stability & Storage

Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. 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 [HRP]

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for HA Tag Antibody [HRP]

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.

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