HA Tag Antibody - BSA Free

Novus Biologicals | Catalog # NB600-362

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Epitope Tag

Cited:

Human, Mouse, Epitope Tag

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Format

BSA Free
View all HA Tag Primary Antibodies »

Product Specifications

Immunogen

This HA Tag Antibody was developed by immunizing goats with HA cleavage site (YPYDVPDYA) conjugated to KLH.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for HA Tag Antibody - BSA Free

Immunohistochemistry: HA Tag Antibody - BSA Free [NB600-362]

Immunohistochemistry: HA Tag Antibody - BSA Free [NB600-362]

HA-Tag-Antibody-Immunohistochemistry-NB600-362-img0011.jpg
Western Blot: HA Tag AntibodyBSA Free [NB600-362]

Western Blot: HA Tag AntibodyBSA Free [NB600-362]

Western Blot: HA Tag Antibody [NB600-362] - Lysates of HEK293T human embryonic kidney cell line transfected with N-terminal HA-tagged MICA, N-terminal HA-tagged LGI-2, mock transfected, C-terminal HA-tagged LRRN5, and C-terminal HA-tagged Trance. PVDF membrane was probed with 1:500 dilution of 1:2500 dilution of goat anti-HA Tag polyclonal (NB600-362, Novus Biologicals), 1:5000 dilution of rabbit anti-HA Tag polyclonal (NB600-363, Novus Biologicals), followed by 1:2000 dilution of the appropriate HRP-conjugated secondary antibody, donkey anti-mouse IgG (HAF018).
Immunocytochemistry/ Immunofluorescence: HA Tag Antibody - BSA Free [NB600-362]

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody - BSA Free [NB600-362]

Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-362] - HA-tagged proteins were detected in immersion fixed HEK293 human embryonic kidney cell line transfected with HA-tagged MGAT2 using 1 ug/mL goat anti-HA Tag polyclonal (NB600-362, Novus Biologicals), 1 ug/mL rabbit anti-HA Tag polyclonal (NB600-363, Novus Biologicals). Cells were stained using the appropriate secondary antibody, donkey anti-mouse IgG-NL557 (NL007) and counterstained with DAPI (blue).
Western Blot: HA Tag AntibodyBSA Free [NB600-362]

Western Blot: HA Tag AntibodyBSA Free [NB600-362]

Western Blot: HA Tag Antibody - BSA Free [NB600-362] - 200, 67, 22, 7, or 2 ng of E. coli whole cell lysateexpressing a multi-tag fusion protein. Antibodies: Affinitypurified, goat anti-HA antibody used for WBat 0.04 ug/ml (1:25,000). Detection: Chemiluminescencewith an exposure time of 10 seconds.
Immunocytochemistry/ Immunofluorescence: HA Tag Antibody - BSA Free [NB600-362]

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody - BSA Free [NB600-362]

Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-362] - HA tagged RFX6 overexpression in HEK293 stained with HA Tag antibody at a dilution of 1:250. ICC/IF image submitted by a verified customer review.
HA Tag Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody - BSA Free [NB600-362] -

SUMO1 conjugates are not localized to amyloid plaques. (a) Sagittal brain sections of 24‐week‐old KI/AD & WT/AD animals were stained using anti‐HA antibodies (red), 6E10 antibodies (green) that label amyloid beta 1–42 among other amyloid beta variants (epitope lies within amino acids 3–8 of amyloid beta) & anti‐MAP2 antibodies (blue). Sections of the hippocampal subiculum are shown. Images are representatives of three independent experiments. Scale bar, 25 μm. (b) Anti‐HA signal intensity in amyloid plaques & in cell nuclei was quantified using ImageJ (N = 3, ***significance between WT/AD & KI/AD, p = .0007 in Student's t test) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29633471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
HA Tag Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody - BSA Free [NB600-362] -

SUMO1 conjugates remain nuclear during increased amyloid burden. (a) Western blot analysis of subcellular fractions of 36‐week‐old KI/WT & KI/AD mouse brain using anti‐HA antibody (upper two panels) & antibodies to GluN1 (a marker of the postsynaptic compartment) & Synaptophysin (a marker of the presynaptic compartment) to validate the fractionation procedure (lower two panels). H, homogenate; P1, nuclear pellet; S1, supernatant after P1 sedimentation; P2, crude synaptosomal pellet; S2, supernatant after P2 sedimentation; LP1, lysed synaptosomal membranes; LS1, supernatant after LP1 sedimentation; LP2, pellet after LS1 sedimentation, SPM, synaptic plasma membranes. Bracket indicates the anti‐HA signal representing SUMOylation, arrow indicates RanGAP1, stars indicate nonspecific signal detected by the anti‐HA antibody. (b) Brain sagittal sections of aged (36 weeks old) KI/AD (left panels) & WT/AD (right panels) mice were immunostained using antibodies directed against HA (red, labels HA‐HA conjugates), MAP2 (green, labels neuronal somata & dendrites), & Synapsin1 (Syn1, magenta, labels synapses). The images show triple‐labeled neurons of hippocampal subiculum (b), cortical layer 5 (c), & proximal apical dendrite from hippocampal CA3 (d). The white line shows the orientation of the line‐scan used to generate the image stack shown in the bottom side view. Scale bar: 10 μm. Note that the anti‐HA immunosignal is mainly located in neuronal nuclei, only background staining is observed in WT/AD mice. Little anti‐HA signal is observed along MAP2‐positive structures & does not colocalize with Synapsin1 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29633471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for HA Tag Antibody - BSA Free

Application
Recommended Usage

ELISA

1:1000 - 1:3000

Immunocytochemistry/ Immunofluorescence

1:100 - 1:400

Immunoprecipitation

2 - 10 ug / mg lysate

Western Blot

1:10000 - 1:25000
Application Notes
ICC/IF reactivity reported in scientific literature (PMID: 23050017).

Reviewed Applications

Read 3 reviews rated 4.3 using NB600-362 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

1.0 mg/ml

Shipping

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

Stability & Storage

Store at 4C. Do not freeze.

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 - BSA Free

Certificate of Analysis

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

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Product Specific Notices for HA Tag Antibody - BSA Free

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.

Related Research Areas

Infectious Diseases

Citations for HA Tag Antibody - BSA Free

Customer Reviews for HA Tag Antibody - BSA Free (3)

4.3 out of 5
3 Customer Ratings
5 Stars
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4 Stars
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • HA Tag Antibody
    Name: Anni Laitinen
    Application: Western Blot
    Sample Tested: HEK 293T cells
    Species: Human
    Verified Customer | Posted 04/28/2017
    First lane is the negative control (untransfected cells) Second lane is HA-tagged WT-RFX6 of 140 kDa Third lane is HA-tagged mutant RFX6, which is a truncated protein of 70 kDa antibody dilution is 1:2000 exposure time is 1 second
    HA Tag Antibody - BSA Free NB600-362
  • HA Tag Antibody
    Name: Anni Laitinen
    Application: Immunocytochemistry
    Sample Tested: 293t HEK and U2OS cells
    Species: Human
    Verified Customer | Posted 04/28/2017
    HA tagged RFX6 overexpression in HEK293 stained at a dilution of 1:250
    HA Tag Antibody - BSA Free NB600-362
  • Name: Lyubov Popova
    Application: Western Blot
    Sample Tested: Hela, Cos whole cell lysates, serum after expression in Baculovirus system
    Species: Other
    Verified Customer | Posted 03/04/2013
    HA Tag Antibody - BSA Free NB600-362

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for HA Tag Antibody - BSA Free

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  • Q: 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?

    A:

    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.

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