HA Tag Antibody - BSA Free

Novus Biologicals | Catalog # NB600-363

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Epitope Tag

Cited:

Human, Mouse, Rat, Epitope Tag, Non-species specific, Yeast

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Western Blot, Block/Neutralize, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chemotaxis, Gel Supershift Assay, Super-Resolution Microscopy, SDS-Page

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all HA Tag Primary Antibodies »

Product Specifications

Immunogen

This HA Tag Antibody was developed by immunizing rabbits with HA cleavage site (YPYDVPDYA) conjugated to KLH. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for HA Tag Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody [NB600-363]

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody [NB600-363]

Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-363] - Detection of HA-tagged Tubulin in CHO cells transfected with mutant beta-tubulin cDNA encoding an HA epitope tag. Photo contributed by R. Bhattacharya and F. Cabral, University of Texas Medical School at Houston.
Western Blot: HA Tag Antibody [NB600-363]

Western Blot: HA Tag Antibody [NB600-363]

Western Blot: HA Tag Antibody [NB600-363] - 200, 100, or 50 ng of E. coli whole cell lysate expressing amulti-tag fusion protein.Antibody: Affinity purified rabbitanti-HA antibody used for WB at 0.04 ug/ml(1:25,000). Detection: Chemiluminescence with anexposure time of 3 seconds.
Western Blot: HA Tag Antibody [NB600-363]

Western Blot: HA Tag Antibody [NB600-363]

HA-Tag-Antibody-Western-Blot-NB600-363-img0018.jpg
Western Blot: HA Tag Antibody [NB600-363]

Western Blot: HA Tag Antibody [NB600-363]

Western Blot: HA Tag Antibody [NB600-363] - ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors and pEGFP, cell lysates were probed with antibodies against the HA tag and EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, and 1200 ng. NT, non-transfected cells. Selection-independent generation of gene knockout mouse embryonic stem cells using zinc-finger nucleases. PLoS One (2011)
Immunocytochemistry/ Immunofluorescence: HA Tag Antibody [NB600-363]

Immunocytochemistry/ Immunofluorescence: HA Tag Antibody [NB600-363]

Immunocytochemistry/Immunofluorescence: HA Tag Antibody [NB600-363] - 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).
HA Tag Antibody

Western Blot: HA Tag Antibody [NB600-363] -

Western Blot: HA Tag Antibody [NB600-363] - Gene knockout in U2OS.693 cells.(A) Schematic of ZFN-mediated knockout. A ZFN pair (ZFNR & ZFNL) designed to target position 502 in the EGFP gene (E502) creates a DNA double-strand break that is sealed by the error-prone non-homologous end-joining (NHEJ) pathway & hence leads to disruption of the coding sequence. (B) Dose-dependent gene disruption in U2OS.693 cells. U2OS.693 cells that stably express a destabilized EGFP were transfected with increasing amounts of E502-specific ZFN expression vectors (75–1200 ng). The percentage of EGFP-negative cells was determined 6 days post-transfection by flow cytometry (n = 3; indicated is average & standard deviation). E502-WT, ZFN with wild-type FokI domain; E502-OH, ZFN with obligate heterodimeric FokI domain. (C) ZFN expression levels. After co-transfection of HEK293T cells with ZFN expression vectors & pEGFP, cell lysates were probed with antibodies against the HA tag & EGFP. Amount of transfected ZFN plasmids was 75 ng, 300 ng, & 1200 ng. NT, non-transfected cells. (D) Kinetics of EGFP knockout. The graph displays the percentage of EGFP-negative cells (see B) from day 1 to day 6 post-transfection for two vector amounts (n = 3; indicated is average & standard deviation). WT, E502-WT; OH, E502-OH. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22194948), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for HA Tag Antibody - BSA Free

Application
Recommended Usage

ELISA

1:100-1:2000

Immunocytochemistry/ Immunofluorescence

1:100-1:400

Immunohistochemistry

1:100-1:400

Immunohistochemistry-Paraffin

1:100-1:400

Immunoprecipitation

1-4 ug/mg lysate

Western Blot

1:1000-1:10000
Application Notes
Use in Gel Super Shift Assays reported in scientific literature (PMID:34289349). Use in ChIP reported in (PMID: 30659200).. Use in IHC reported in (PMID: 22880041).. Use in FLOW reported in scientific literature (Pryce R et al).. Knockout validation (PMID: 32354171).. Recommended dilution for coating ELISA plates is 1:100 - 1:500.

Reviewed Applications

Read 3 reviews rated 4.3 using NB600-363 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.09% Sodium Azide

Concentration

0.1 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)

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3 Customer Ratings
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • HA Tag Antibody
    Name: Tanya Kutyavin
    Application: Simple Western
    Sample Tested: Recombinant proteins
    Species: Human
    Verified Customer | Posted 06/08/2018
    HA Tag Antibody - BSA Free NB600-363
  • Name: Filipa Franquinho
    Application: Western Blot
    Sample Tested: Hek 293T cell lysate
    Species: Human
    Verified Customer | Posted 05/04/2013
  • Name: Maria Andreou
    Application: Western Blot
    Sample Tested: Xenopus laevis inejected embryo Lysates, Sample Amount: 30ug
    Species: Other
    Verified Customer | Posted 09/26/2011
    HA Tag Antibody - BSA Free NB600-363

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Protocols

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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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