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HA Tag Antibody (543851)

R&D Systems | Catalog # MAB060

R&D Systems

Key Product Details

Species Reactivity

Validated:

Multi-Species

Cited:

Human, Mouse

Applications

Validated:

Western Blot

Cited:

Neutralization, Bioassay, Cross-linking, Functional Assay

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 543851
View all HA Tag Primary Antibodies »

Product Specifications

Immunogen

HA Tag Peptide
Accession # ABB51961

Specificity

Specifically detects N-terminal HA-tagged proteins with a free N-terminus.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for HA Tag Antibody (543851)

Detection of N-terminal HA-tag antibody by Western Blot.

Detection of N‑terminal HA-tag by Western Blot.

Western blot shows Recombinant Human APRIL/TNFSF13 (Catalog # 5860-AP, 5 ng/lane). PVDF membrane was probed with 1 µg/mL of Mouse Anti-HA Tag Peptide Antibody Monoclonal Antibody (Catalog # MAB060) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for N-terminal HA-tagged Recombinant Human APRIL/ TNFSF13 at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of HA Tag by Flow Cytometry

Detection of HA Tag by Flow Cytometry

Cell survival and IgG secretion of encapsulated mouse bone marrow plasma cells over time.(a) Cell viability of encapsulated and non-encapsulated cells after incubation in culture medium for different time periods. Viability was analyzed by flow cytometry (gated on single cells or single beads/CD138+/live; gating strategy in Supplementary Fig. 2). (b) Detection of IgG in the culture supernatant of encapsulated (red) and nonencapsulated cells (dark gray) by ELISA at different time points. The dashed line indicates the sensitivity of the assay (absorbance corresponding to 1.56 ng/mL IgG). The solid lines denote the mean of two technical replicates. (c) IgG1 secretion of encapsulated and non-encapsulated cells over time was analyzed by flow cytometry. To determine the extent of cross-contamination between beads, the IgG1 signal of plasma cells (CD138+) was compared to the signal of empty beads or cells that lack the plasma cell marker (CD138−; gating strategy in Supplementary Fig. 3). The numbers in the top right corner indicate the percentage of IgG+ events in the CD138+ (red) and CD138− (blue) populations (IgG+ gate indicated by a gray line). The number in the bottom right corner indicates the percentage of ‘true positives’ (percentage of IgG+ events originating from CD138+ based on the sum of IgG+ events from both CD138+ and CD138− populations). The plots show a minimum of 10,019 (CD138−) and 2,533 (CD138+) events at 2% contour level. (d) Time course of ASC IgG secretion and background based on c. Left axis: percentage of IgG1-positive events in the CD138+ (red diamonds) and CD138− (blue diamonds) populations for each analyzed time point. Right axis (gray diamonds): percentage of events in the IgG1 gate that originate from CD138+ and not from CD138−.Source data Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41587-024-02346-5), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for HA Tag Antibody (543851)

Application
Recommended Usage

Western Blot

1 µg/mL
Sample: N-terminal HA-tagged Recombinant Human APRIL/TNFSF13 (Catalog # 5860-AP)

Reviewed Applications

Read 8 reviews rated 4.8 using MAB060 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

HA

UniProt

ABB51961

Additional HA Tag Products

Product Documents for HA Tag Antibody (543851)

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for HA Tag Antibody (543851)

For research use only

Related Research Areas

Infectious Diseases

Citations for HA Tag Antibody (543851)

Customer Reviews for HA Tag Antibody (543851) (8)

4.8 out of 5
8 Customer Ratings
5 Stars
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4 Stars
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3 Stars
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Showing  1 - 5 of 8 reviews Showing All
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  • HA Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 11/04/2021
    HA Tag Antibody (543851) MAB060
  • HA Tag Antibody
    Name: Lucia Sereni
    Application: Cell Proliferation
    Sample Tested: B cells
    Species: Mouse
    Verified Customer | Posted 01/05/2021
    HA Tag Antibody (543851) MAB060
  • HA Tag Antibody
    Name: Anonymous
    Application: Cell Culture
    Sample Tested: B cells
    Species: Mouse
    Verified Customer | Posted 10/22/2020
    This product has been used in combination with murine CD40L-HaTag protein to culture primary murine B cells.
    HA Tag Antibody (543851) MAB060
  • HA Tag Antibody
    Name: Kurt Shanebeck
    Application: Functional Assay
    Sample Tested: Recombinant protein
    Species: Human
    Verified Customer | Posted 12/06/2017
  • HA Tag Antibody
    Name: Shub Poddar
    Application: Flow Cytometry
    Sample Tested: Jurkat human acute T cell leukemia cell line
    Species: Human
    Verified Customer | Posted 08/23/2017
  • HA Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293-EBNA human EBV-expressing embryonic kidney cell line
    Species: Mouse
    Verified Customer | Posted 07/23/2017
    HA Tag Antibody (543851) MAB060
  • HA Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 07/21/2017
    HA Tag Antibody (543851) MAB060
  • HA Tag Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: 293-EBNA human EBV-expressing embryonic kidney cell line
    Species: Mouse
    Verified Customer | Posted 07/20/2017
    HA Tag Antibody (543851) MAB060

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