Overview

  • Product name
    Anti-HA tag antibody [HA.C5]
    See all HA tag primary antibodies
  • Description
    Mouse monoclonal [HA.C5] to HA tag
  • Host species
    Mouse
  • Tested applications
    Suitable for: ChIP/Chip, WB, ICC, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Synthetic peptide from influenza hemagglutinin epitope:

    YPYDVPDYA

    conjugated to KLH.

  • General notes

    This product was changed from ascites to tissue culture supernatant on 5th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

    See other anti-mouse secondary antibodies that can be used with this antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.05% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    Purified from TCS
  • Clonality
    Monoclonal
  • Clone number
    HA.C5
  • Isotype
    IgG3
  • 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 Abreviews Notes
ChIP/Chip Use at an assay dependent concentration. PubMed: 19581286
WB 1/1000.
ICC 1/200.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Relevance
    Human 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

Images

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

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

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : 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

    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

  • 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
    All lanes : HRP conjugated Goat anti-mouse IgG (H+L)

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 10 seconds


    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

References

This product has been referenced in:
  • Li Z  et al. APC-Cdh1 Regulates Neuronal Apoptosis Through Modulating Glycolysis and Pentose-Phosphate Pathway After Oxygen-Glucose Deprivation and Reperfusion. Cell Mol Neurobiol 39:123-135 (2019). Read more (PubMed: 30460429) »
  • Sun G  et al. Hsc70 Interacts with ß4GalT5 to Regulate the Growth of Gliomas. Neuromolecular Med 21:33-41 (2019). Read more (PubMed: 30607818) »
See all 88 Publications for this product

Customer reviews and Q&As

11-20 of 20 Abreviews or Q&A

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

Abcam user community

Verified customer

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

Dr. Harendra Chahar

Verified customer

Submitted Sep 10 2012

Answer

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: ab9110) and the new order number is 961284. I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product. Good luck with your research!

Read More
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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

Submitted Dec 09 2010

Application
Western blot
Sample
Mouse Recombinant protein (test expression)
Loading amount
50 µg
Specification
test expression
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted May 17 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Renal Carcinoma Cell Line)
Specification
Renal Carcinoma Cell Line
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 4%

Abcam user community

Verified customer

Submitted Aug 23 2006

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (osteosarcoma U2OS)
Specification
osteosarcoma U2OS
Fixative
Paraformaldehyde
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1

Abcam user community

Verified customer

Submitted Jul 27 2006

Application
Western blot
Sample
Human Cell lysate - whole cell (Renal Carcinoma Cell Line)
Loading amount
50 µg
Specification
Renal Carcinoma Cell Line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4%

Abcam user community

Verified customer

Submitted Jul 26 2006

Answer

Thank you for your enquiry. NChIP involves the preparation of native, nuclease digested chromatin followed by the immunoprecipitation of chromatin using appropriate antibodies: We have an excellent protocol for both the preparation of nuclease digested chromatin: http://ops.abcam.com/index.html?pageconfig=resource&rid=31&pid=5 and the immunoprecipitation of chromatin: http://ops.abcam.com/index.html?pageconfig=view_protocol&pid=293. Both of these protocols were developed by Laura O'Neill and Bryan Turner. They have published their NChiP protocol in the following publication: O'Neill LP, Turner BM. (2003) Immunoprecipitation of native chromatin: NChIP. Methods. 31(1):76-82. PMID: 12893176 Please forgive me if I am incorrect but given that you are using crosslinking in your detection of transcription factor interaction with genomic DNA your approach is more akin to XChIP. I would recommend an epitope tag that has been validated for its use by chromatin immunoprecipitation. We have an excellent antibody in the form of ab9110 HA, tag antibody - ChIP Grade. This has been shown to be capable of precipitating chromatin under cross linking conditions. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

11-20 of 20 Abreviews or Q&A

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