Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with N-terminal HA-tagged LGI-2, Internal Sequence HA-tagged CIGALT, and C-terminal HA-tagged FBLN. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-HA Peptide Monoclonal Antibody (Catalog # MAB6875) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for HA-tagged proteins at approximately 60, 37, and 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

HA Tag in HEK293 Human Cell Line.

HA Tag was detected in immersion fixed HEK293 human embryonic kidney cell line using Mouse Anti-HA Tag Monoclonal Antibody (Catalog # MAB6875) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of HA in HEK293 Human Cell Line Transfected with HA-tagged Proteins by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with HA-tagged proteins was stained with Mouse Anti-HA Monoclonal Antibody (Catalog # MAB6875, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

Applications for HA Tag Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HEK293 human embryonic kidney cell line

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: HEK293 human embryonic kidney cell line transfected with Hemagglutinin/HA-tagged proteins fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

Western Blot

0.5 µg/mL
Sample: HEK293 human embryonic kidney cell line transfected with N-terminal HA-tagged LGI-2, Internal Sequence HA-tagged CIGALT, and C-terminal HA-tagged FBLN

Reviewed Applications

Read 4 reviews rated 3.3 using MAB6875 in the following applications:

Immunocytochemistry/Immunofluorescence (1 Review)
Immunohistochemistry (1 Review)
Western Blot (2 Reviews)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

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

Entrez Gene IDs

23308115 (Influenza A Virus H1N1); 3655151 (Influenza A Virus H3N2)

Gene Symbol

UniProt

ABB51961

Additional HA Tag Products

Product Documents for HA Tag Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for HA Tag Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

HA Tag Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB6875

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

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Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for HA Tag Antibody

Detection of HA-tagged proteins by Western Blot.