HA Tag Antibody - BSA Free

Immunohistochemistry: HA Tag Antibody [NB600-362] - SUMO1 conjugates are not localized to amyloid plaques. (a) Sagittal brain sections of 24-week-old KI/AD and 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) and anti-MAP2 antibodies (blue). Sections of the hippocampal subiculum are shown. Images are representatives of three independent experiments. Scale bar, 25 um. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/acel.12760) licensed under a CC-BY license.

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 [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 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 [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.

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 (//pubmed.ncbi.nlm.nih.gov/29633471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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 (//pubmed.ncbi.nlm.nih.gov/29633471), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: ET
• Applications: WB, ELISA, ICC/IF, IHC, IP
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

HA Tag Antibody - BSA Free Summary

• Immunogen: This HA Tag Antibody was developed by immunizing goats with HA cleavage site (YPYDVPDYA) conjugated to KLH.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:1000 - 1:3000
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:400
  • Immunohistochemistry
  • Immunoprecipitation 2 - 10 ug / mg lysate
  • Western Blot 1:10000 - 1:25000
• Application Notes: ICC/IF reactivity reported in scientific literature (PMID: 23050017).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS
• Preservative: 0.09% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for HA Tag Antibody - BSA Free

• HA Epitope Tag
• HA Tag
• HA
• Hemagglutinin

Background

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

Limitations

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.

Publications for HA Tag Antibody (NB600-362)(42)

Publications using NB600-362

• Wünkhaus D, Tang R, Nyame K et Al. TRPML1 activation ameliorates lysosomal phenotypes in CLN3 deficient retinal pigment epithelial cells Sci Rep 2024-07-30 [PMID: 39080379]
• Murata D, Roy S, Lutsenko S, Iijima M et Al. Slc25a3-dependent copper transport controls flickering-induced Opa1 processing for mitochondrial safeguard Dev Cell 2024-07-10 [PMID: 38986607]
• Timothy M O'Shea, Yan Ao, Shinong Wang, Yilong Ren, Amy L Cheng, Riki Kawaguchi, Zechuan Shi, Vivek Swarup, Michael V Sofroniew Derivation and transcriptional reprogramming of border-forming wound repair astrocytes after spinal cord injury or stroke in mice. Nature neuroscience 2024-06-21 [PMID: 38907165]
• Hellweg L, Pfeifer M, Tarnawski M, Thing-Teoh S et Al. AspSnFR: A genetically encoded biosensor for real-time monitoring of aspartate in live cells Cell Chem Biol 2024-05-28 [PMID: 38806058]
• Lin MC, Kuo WH, Chen SY et Al. Ago2/CAV1 interaction potentiates metastasis via controlling Ago2 localization and miRNA action EMBO Rep 2024-04-22 [PMID: 38649663]
• Carlene L Zindl, C Garrett Wilson, Awalpreet S Chadha, Lennard W Duck, Baiyi Cai, Stacey N Harbour, Yoshiko Nagaoka-Kamata, Robin D Hatton, Min Gao, David A Figge, Casey T Weaver Distal colonocytes targeted by C. rodentium recruit T-cell help for barrier defence. Nature 2024-04-10 [PMID: 38600382]
• McCarron KR, Elcocks H, Mortiboys H et Al. The Parkinson's disease related mutant VPS35 (D620N) amplifies the LRRK2 response to endolysosomal stress Biochem J 2024-02-21 [PMID: 38299383]
• Adewumi HO, Berniac GI, McCarthy EA et Al. Ischemic and hemorrhagic stroke lesion environments differentially alter the glia repair potential of neural progenitor cell and immature astrocyte grafts Exp Neurol 2024-01-18 [PMID: 38244885]
• Lee B, Kim YH, Lee W et al. USP13 deubiquitinates p62/SQSTM1 to induce autophagy and Nrf2 release for activating antioxidant response genes Free radical biology & medicine 2023-11-01 [PMID: 37776917] (WB, BA)
• Padgett L, Shinkle M, Rosario S et al. Deoxyhypusine synthase (DHPS) mutations alter the post-translational modification of eukaryotic translation factor 5A (eIF5A) resulting in impaired human and mouse neural homeostasis Human Genetics and Genomics Advances 2023-05-01 [PMID: 37333770] (Western Blot, Epitope Tag)
• Yan Y, Gauthier MA, Malik A et al. The NOTCH-RIPK4-IRF6-ELOVL4 Axis Suppresses Squamous Cell Carcinoma Cancers (Basel) 2023-01-25 [PMID: 36765696]
• Pfeiffer A, Petersen JD, Falduto GH et al. Selective immunocapture reveals neoplastic human mast cells secrete distinct microvesicle- and exosome-like populations of KIT-containing extracellular vesicles Journal of extracellular vesicles 2022-09-01 [PMID: 36239715] (IP)
• Wei G, Iqbal N, Courouble VV et al. Prion-like low complexity regions enable avid virus-host interactions during HIV-1 infection Nature Communications 2022-10-06 [PMID: 36202818]
• O'Shea TM, Ao Y, Wang S et al. Lesion environments direct transplanted neural progenitors towards a wound repair astroglial phenotype in mice Nature Communications 2022-09-28 [PMID: 36171203]
• Park H, Seo SK, Sim JR et al. TMED3 Complex Mediates ER Stress-Associated Secretion of CFTR, Pendrin, and SARS-CoV-2 Spike Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2022-06-24 [PMID: 35748162]
• Burda JE, O'Shea TM, Ao Y et al. Divergent transcriptional regulation of astrocyte reactivity across disorders Nature 2022-05-25 [PMID: 35614216] (IHC-Fr)
• Adilardi RS, Dernburg AF. Robust, versatile DNA FISH probes for chromosome-specific repeats in Caenorhabditis elegans and Pristionchus pacificus G3 Genes|Genomes|Genetics 2022-07-06 [PMID: 35567480]
• Zaug AJ, Lim CJ, Olson CL et al. CST does not evict elongating telomerase but prevents initiation by ssDNA binding Nucleic Acids Research 2021-11-18 [PMID: 34718732] (Western Blot)
• Rillo-Bohn R, Adilardi R, Avsaroglu B et al. Analysis of meiosis in Pristionchus pacificus reveals plasticity in homolog pairing and synapsis within the nematode lineage Elife 2021-08-24 [PMID: 34427184]
• Natalia Malek, Aleksandra Michrowska, Ewa Mazurkiewicz, Ewa Mrówczyńska, Paweł Mackiewicz, Antonina J. Mazur The origin of the expressed retrotransposed gene ACTBL2 and its influence on human melanoma cells’ motility and focal adhesion formation Scientific Reports 2021-02-08 [PMID: 33558623]
• Kaneko Y, Shimoda K, Ayala R et al. p97 and p47 function in membrane tethering in cooperation with FTCD during mitotic Golgi reassembly The EMBO journal 2021-02-08 [PMID: 33555040]
• Adachi Y, Kato T, Yamada T et al. Drp1 Tubulates the ER in a GTPase-Independent Manner Mol Cell 2020-11-19 [PMID: 33152269]
• Son SM, Park SJ, Stamatakou E et al. Leucine regulates autophagy via acetylation of the mTORC1 component raptor Nat Commun 2020-06-19 [PMID: 32561715] (WB, Mouse)
• Zeng J, Xu H, Fan PZ et al. Kaempferol blocks neutrophil extracellular traps formation and reduces tumour metastasis by inhibiting ROS-PAD4 pathway Journal of Cellular and Molecular Medicine 2020-07-01 [PMID: 32427405]
• Ganapathi M, Padgett LR, Yamada K et al. Recessive Rare Variants in Deoxyhypusine Synthase, an Enzyme Involved in the Synthesis of Hypusine, Are Associated with a Neurodevelopmental Disorder Am J Hum Genet 2019-01-22 [PMID: 30661771] (WB)
• Son S. M, Park S. J, et al. Leucine Signals to mTORC1 via Its Metabolite Acetyl-Coenzyme A. Cell Metab 2019-01-08 [PMID: 30197302] (WB, Human)
• Uchida S, Horie R, Sato H et al. Possible role of the Nipah virus V protein in the regulation of the interferon beta induction by interacting with UBX domain-containing protein1 Sci Rep 2018-05-16 [PMID: 29769705] (WB, Human)
• Nguyen JT, Ray C, Fox AL et al. Mammalian EAK-7 activates alternative mTOR signaling to regulate cell proliferation and migration Sci Adv 2018-05-01 [PMID: 29750193] (IP)
• Hossain D, Ferreira Barbosa JA, Cohen EA, Tsang WY. HIV-1 Vpr hijacks EDD-DYRK2-DDB1(DCAF1) to disrupt centrosome homeostasis. J Biol Chem 2018-06-15 [PMID: 29724823]
• Cartwright Ian M, Liu Xinjian, Zhou Min et al. Essential roles of Caspase-3 in facilitating Myc-induced genetic instability and carcinogenesis. ELife 2017-01-01 [PMID: 28691902] (WB, Human)
• Ain S, Yoon SY, Zhu L et al. Arf4 Determines Dentate Gyrus-Mediated Pattern Separation by Regulating Dendritic Spine Development PLoS One 2012-01-01 [PMID: 23050017] (ICC/IF, Mouse)
• Peterson AR Glutamate Uptake Regulation as a Therapeutic Strategy in Epilepsy Thesis 2022-01-01 (IHC-Fr, WB)
• Itoh K, Murata D, Kato T et al. Brain-specific Drp1 regulates postsynaptic endocytosis and dendrite formation independently of mitochondrial division Elife 2019-10-11 [PMID: 31603426] (ICC/IF)
• Nguyen J T, Haidar F S et al. mEAK-7 Forms an Alternative mTOR Complex with DNA-PKcs in Human Cancer. iScience 2019-07-26 [PMID: 31288154] (IP)
• Wollenberg AL, O'Shea TM, Kim JH et al. Injectable polypeptide hydrogels via methionine modification for neural stem cell delivery Biomaterials 2018-09-01 [PMID: 29657091]
• Popova L, Smith K, West AH et al. Immunodominance of Antigenic Site B over Site A of Hemagglutinin of Recent H3N2 Influenza Viruses plos One 2012-01-01 [PMID: 22848649] (WB)
• Rillo-Bohn R, Adilardi R, Avsaroglu B et al. Analysis of meiosis in Pristionchus pacificus reveals plasticity in homolog pairing and synapsis within the nematode lineage bioRxiv
• Sharma M, Giridharan SS, Rahajeng J et al. MICAL-L1 links EHD1 to tubular recycling endosomes and regulates receptor recycling. Mol Biol Cell 2009-12-01 [PMID: 19864458]
• Abedini MR, Muller EJ, Brun J et al. Cisplatin induces p53-dependent FLICE-like inhibitory protein ubiquitination in ovarian cancer cells. Cancer Res 2008-06-01 [PMID: 18559494]
• Broadbent D, McEwan C, Tsang T et al. The formation of ubiquitin rich condensates triggers recruitment of the ATG9A lipid transfer complex to initiate basal autophagy bioRxiv 2023-11-30
• Adewumi H, Berniac G, McCarthy E, O’Shea T Ischemic and hemorrhagic stroke lesion environments differentially alter the glia repair potential of neural progenitor cell and immature astrocyte grafts bioRxiv 2023-08-17 (ICC/IF)
• Flannery, A et al. Topological Characterization of the c, c' c Subunits of the Vacuolar ATPase from the Yeast Saccharomyces cerevisiae. The Journal of Biological Chemistry 279(38): 39856-39862. 2004-01-01 [PMID: 15252052]

Reviews for HA Tag Antibody (NB600-362) (3)

Average Rating: 4.3

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Other.

reviewed by: Anni Laitinen WB Human 04/28/2017

Summary

• Application: Western Blot
• Sample Tested: HEK 293T cells
• Species: Human

reviewed by: Anni Laitinen ICC Human 04/28/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: 293t HEK and U2OS cells
• Species: Human

reviewed by: Lyubov Popova WB Other 03/04/2013

Summary

• Application: Western Blot
• Sample Tested: Hela, Cos whole cell lysates, serum after expression in Baculovirus system
• Species: Other

FAQs for HA Tag Antibody (NB600-362). (Showing 1 - 1 of 1 FAQs).

1. 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?
   • 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.