Overview

  • Product name
    Anti-HA tag antibody - ChIP Grade
    See all HA tag primary antibodies
  • Description
    Rabbit polyclonal to HA tag - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    ELISA: The anti HA diluted 1:70.000 gave an O.D.=1.0 in a 15 minute reaction against peptide conjugated with a different carrier than used for anti peptide purification. HRP conjugated Goat anti rabbit IgG was used and TMB was the substrate.
  • Tested applications
    Suitable for: ChIP/Chip, IP, ELISA, WB, ICC/IF, ICC, Flow Cyt, ChIPmore details
  • Immunogen

    YPYDVPDYA (influenza hemagglutinin-HA-epitope) conjugated to KLH.

  • Positive control
    • WB: 293FT cells transfected with 15kDa HA tagged Vpr (an HIV1 accessory protein). IP: Nuclear lysate of HEK-293T cells transiently expressing HA-tagged protein. ICC/IF: U-2 cells. Mouse olineu cells. ChIP: Xenopus laevis oocytes were injected with mRNA for HA-tagged human BORIS.
  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit 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: 0.1% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antibodies were immunoaffinity purified using the peptide conjugated to a solid-phase support.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9110 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.
IP Use at an assay dependent concentration.
ELISA 1/200 - 1/500.
WB 1/4000 - 1/10000.
ICC/IF Use a concentration of 1 - 4 µg/ml.
ICC Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

ChIP Use 3 µg for 25 µg of chromatin.

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

  • Arrb2 depends on Gβγ to induce membrane translocation of PKCα

    Xenopus embryos were injected with 500 pg pkcα-gfp RNA and co-injected as indicated above the images. Animal Caps were prepared at stage 10 and immunostained as indicated. Nuclei were stained with Hoechst 33258 (blue). Images show representative results from at least two independent experiments with a minimum of six Animal Caps per experiment. Scale bars: 50 µm.

    The inhibitory effect of Arrb2 MO1 (E) on PKCα-GFP membrane translocation was rescued by (F) co-injection of HA-Gβ and HA-Gγ mRNA (anti-HA (red): F′, merge: F").

    HA was detected with ab9110.

    (After Figure 2 of Seitz et al)

     

  • ab9110 was diluted to 4 µg/mg lysate and incubated with a nuclear lysate of HEK293T cells transiently expressing HA-tagged protein and a Protein A matrix for 2 hours a 23°C to achieve immunoprecipitation.  1000 µg of lysate was present in the input. 

    A HRP-conjugated anti-rabbit HA monoclonal antibody diluted 1/1000 was used for the Western Blot step.

    See Abreview

  • This image was kindly supplied as part of the review submitted by Kasper Fugger. Immunofluorescence staining of U-2 cells expressing HA-tagged protein with ab9110.
  • Xenopus laevis oocytes were injected with mRNA for HA-tagged human BORIS. Chromatin was prepared according to the Abcam X-ChIP protocol. Oocytes were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 20µl of Protein A/G sepharose beads, and 3µg of ab9110 (anti-HA, light blue) or, 3µg of ab18337 (anti-Boris, dark blue). A non-specific antibody was used as a control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).

  • All lanes : Anti-HA tag antibody - ChIP Grade (ab9110) at 1/4000 dilution

    Lane 1 : 15ug untransfected wcl lysate
    Lane 2 : 293FT cells transfected with 15kDa HA tagged Vpr (an HIV1 accessory protein)

    Secondary
    All lanes : HRP conjugated Goat anti-Rabbit

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 5 seconds


    This image is courtesy of an Abreview

    See Abreview

  • ab 9110 at a 1/200 dilution staining the mouse olineu cell line (oligodendrocyte precursor cell) by immunocytochemistry. The antibody was incubated with the cells for 30 minutes and then detected using a Cy5 conjugated goat anti-rabbit antibody.

    This image is courtesy of an Abreview submitted by Katarina Trajkovic on 15 March 2006

    See Abreview

References

This product has been referenced in:
  • Rengachari S  et al. Structural basis of STAT2 recognition by IRF9 reveals molecular insights into ISGF3 function. Proc Natl Acad Sci U S A 115:E601-E609 (2018). Read more (PubMed: 29317535) »
  • Albee LJ  et al. Identification and functional characterization of arginine vasopressin receptor 1A : atypical chemokine receptor 3 heteromers in vascular smooth muscle. Open Biol 8:N/A (2018). Read more (PubMed: 29386406) »
See all 498 Publications for this product

Customer reviews and Q&As

1-10 of 12 Q&A

Question
Answer

Although we have not tested this antibody specifically against proteins expressed in S. cerevisiae, it should recognise the protein in this species.

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Question
Answer

I have to confirm that we did not test ab9110 with multiple HA tag repeats in CHIP but only with a single tag.

But I would expect ab9110 to work also with multiple HA tag repeats.

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Question
Answer

Multiple freeze-thaw cycles may reduce the activity of this antibody. If you are concerned about losing activity by freezing this antibody, it can be stored in 50% glycerol at -20C. This will keep the antibody from freezing while allowing it to be stored at a cold enough temperature for it to remain stable long term.

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Answer

I can confirm that this primary antibody (ab9110) can be used for ELISA. Since it is not labelled it is important to use a compatible secondary antibody for detection which is conjugated to either to an enzyme or to a fluorochrome.

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Answer

Yes, ab9110 is the unconjugated form of ab26228 (biotinylated form).

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Answer

Both antibodies have not yet been tested in any IHC application (neither IHC-P or IHC-Fr).

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Answer

Yes, you are correct, this antiserum was validated using our unique system of HA-tagging BORIS and injecting into Xenopus oocytes. The antiserum has not yet been tested by ChIP in human cells. However, this antiserum was tested by ChIP using the standard Abcam XChIP protocol (https://www.abcam.com/index.html?pageconfig=view_protocol&pid=171). Therefore I would strongly predict that it would be immediately transferable to ChIP in human cells.

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Answer

Under the IF suggested dilution range for this antibody, the 1:40,000 dilution was a typo - it was supposed to be 1:4000. We have received some nice feedback from customers who have used this antibody in IF, and several have mentioned obtaining good results using PFA for fixation.

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Answer

This product will recognize the C-terminus, N-terminus and middle.

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1-10 of 12 Q&A

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