Overview

  • Product nameAnti-HA tag antibody [HA.C5]See all HA tag primary antibodies ...
  • Description
    Mouse monoclonal [HA.C5] to HA tag
  • SpecificityRecognizes HA-tagged proteins overexpressed in cells, including both amino- and carboxy-termini of targeted proteins in transfected mammalian cells.
  • Tested applicationsChIP/Chip, WB, ICC, IP, ICC/IF more details
  • Immunogen

    Synthetic peptide from influenza hemagglutinin epitope: YPYDVPDYA conjugated to KLH.

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.05% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • PurityIgG fraction
  • Clonality Monoclonal
  • Clone numberHA.C5
  • IsotypeIgG3
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab18181 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ChIP/Chip Use at an assay dependent dilution.
WB 1/1000.
ICC 1/200.
IP Use at an assay dependent dilution.
ICC/IF Use at an assay dependent dilution.

Target

  • RelevanceHuman influenza hemagglutinin (HA) is a surface glycoprotein required for the infectivity of the human virus. The HA tag is derived from the HA molecule corresponding to amino acids 98-106 has been extensively used as a general epitope tag in expression vectors. Many recombinant proteins have been engineered to express the HA tag, which does not appear to interfere with the bioactivity or the biodistribution of the recombinant protein. This tag facilitates the detection, isolation, and purification of the proteins.
  • Alternative names
    • HA epitope tag antibody
    • HA1 antibody
    • HA2 antibody
    • hemagglutinin antibody
    • Hemagglutinin HA1 chain antibody
    • Hemagglutinin HA2 chain antibody
    see all

Anti-HA tag antibody [HA.C5] images

  • All lanes : Anti-HA tag antibody [HA.C5] (ab18181) at 1/1000 dilution

    Lane 1 : HEK293 whole cell lysate - transfected
    Lane 2 : HEK293 whole cell lysate - transfected
    Lane 3 : HEK293 whole cell lysate - untransfected

    Lysates/proteins at 30 µg per lane.

    Secondary
    IRDye® 800CW Goat anti-mouse IgG polyclonal at 1/10000 dilution

    Performed under reducing conditions.

    Observed band size : 85 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Western blot using ab18181 of 293 cells transfected with HA-tagged vector(2) and untransfected control (1). Western blot using ab18181 of 293 cells transfected with HA-tagged vector(2) and untransfected control (1).
  • Immunofluorescence using ab18181 staining a HA-tag fusion protein (transcription factor) in a stable expressing cell line (right hand panel) and control cell line (left hand panel).
  • All lanes : Anti-HA tag antibody [HA.C5] (ab18181) at 1/2000 dilution

    Lane 1 : WCE from cell line transfected for HA-tagged protein
    Lane 2 : WCE from a cell line transfected with empty vector

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP conjugated Goat anti-mouse IgG (H+L)
    developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 10 seconds

    This image is courtesy of an anonymous Abreview

    Incubation with the primary antibody was carried out at 4°C overnight, whilst the secondary antibody was incubated for 1 hour at room temperature.

    See Abreview

  • ab18181 staining HA tag (green) in HeLa cells by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 3% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/1000 in diluent) for 1 hour at 22°C. A FITC-conjugated goat anti-mouse polyclonal IgG (1/1000) was used as the secondary antibody. Nuclei were stained with DAPI (blue).

    See Abreview

References for Anti-HA tag antibody [HA.C5] (ab18181)

This product has been referenced in:
  • van Kasteren PB  et al. Deubiquitinase function of arterivirus papain-like protease 2 suppresses the innate immune response in infected host cells. Proc Natl Acad Sci U S A 110:E838-47 (2013). WB . Read more (PubMed: 23401522) »
  • Lang UE  et al. GSK3ß phosphorylation of the KLF6 tumor suppressor promotes its transactivation of p21. Oncogene 32:4557-64 (2013). Read more (PubMed: 23085750) »

See all 16 Publications for this product

Product Wall

Application Immunoprecipitation
Immuno-precipitation step Protein G
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Treatment untransfected/ transfected ha-tagged protein
Total protein in input 100 µg
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Submitted Nov 27 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Blocking step Odyssey block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
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Submitted Nov 25 2013

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (CHO cells)
Total protein in input 500 µg
Specification CHO cells
Immuno-precipitation step Protein A/G
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Submitted Feb 26 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HepG2)
Specification HepG2
Fixative Paraformaldehyde
Permeabilization Yes - Saponin 0.1%
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 19°C
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Submitted Feb 12 2013

Application Western blot
Sample Human Cell lysate - whole cell (293T)
Loading amount 15 µg
Specification 293T
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 8% · Temperature: 20°C
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Mr. Dongil Kim

Verified customer

Submitted Jan 24 2013

Application Western blot
Sample Human Cell lysate - whole cell (HA tagged Viarl protein expressed in Hela)
Loading amount 20 µg
Specification HA tagged Viarl protein expressed in Hela
Gel Running Conditions Reduced Denaturing (4-10%)
Blocking step BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Sep 11 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% tritonx100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
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Dr. Harendra Chahar

Verified customer

Submitted Sep 10 2012

Thank you for getting back to me. I am sorry that my previous e-mail has not reached you. This is to let you know that I have placed a new order for you - for one vial of ab18181 as a free of charge replacement (exchange for the original item: a...

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Application Western blot
Sample Human Cell lysate - whole cell (neuroblastoma cell)
Loading amount 25 µg
Specification neuroblastoma cell
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Feb 04 2011

Application Western blot
Sample Chinese Hamster Cell lysate - whole cell (CHO-cells)
Loading amount 200000 cells
Specification CHO-cells
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Dec 09 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"