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.

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

  • 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

  • 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).

  • 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 447 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (Co-IP from mouse NSCs)
Total protein in input
500 µg
Immuno-precipitation step
Protein A/G
Specification
Co-IP from mouse NSCs
Username

Abcam user community

Verified customer

Submitted Aug 01 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse neural stem cell)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
40 µg
Specification
mouse neural stem cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jun 12 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 cells)
Gel Running Conditions
Reduced Denaturing (4-12% gradient)
Loading amount
15 µg
Specification
HEK293 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Rebecca Fu

Verified customer

Submitted Apr 17 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Username

Dr. Jan Tuma

Verified customer

Submitted Sep 21 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (alveolar type II cell transfected with a HA-Tagged)
Permeabilization
Yes - Saponin
Specification
alveolar type II cell transfected with a HA-Tagged
Blocking step
Serum as blocking agent for 5 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 18 2017

Application
ChIP
Sample
Mouse Cell lysate - nuclear (mouse embrynic stem cells)
Negative control
Promoters Sdha and b-Act
Specification
mouse embrynic stem cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldhyde 0.75%
Positive control
Promoters of Cbx8 and Hmox1
Username

Abcam user community

Verified customer

Submitted Apr 21 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.1% Triton X-100/0.02% SDS
Specification
HeLa
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
3% paraformaldehyde, 0.02% glutaraldehyde
Username

Abcam user community

Verified customer

Submitted Jan 25 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Whole cell lysate resolved on a 12% SDS PAGE gel)
Gel Running Conditions
Reduced Denaturing (12% SDS gel)
Loading amount
25 µg
Treatment
HEK293T cells transfected with HA-Rad21 plasmid for overexpression
Specification
Whole cell lysate resolved on a 12% SDS PAGE gel
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 19 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Embryonic kidney fibroblast cells)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
1e+006 cells
Specification
Embryonic kidney fibroblast cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Apr 01 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Cell (spinal cord)
Permeabilization
Yes - PBS/gelatin/triton/serum
Specification
spinal cord
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 14 2015

1-10 of 63 Abreviews or Q&A

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