Overview

  • Product name
    Anti-HA tag antibody - ChIP Grade
    See all HA tag primary antibodies
  • Description
    Rabbit polyclonal to HA tag - ChIP Grade
  • 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.

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
    HRP conjugated Goat anti-Rabbit
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 5 secondsThis 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:
  • He Y  et al. A Pap1-Oxs1 signaling pathway for disulfide stress in Schizosaccharomyces pombe. Nucleic Acids Res 45:106-114 (2017). IP . Read more (PubMed: 27664222) »
  • Jung J  et al. Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator. Elife 6:N/A (2017). WB, ICC/IF . Read more (PubMed: 28195531) »

See all 385 Publications for this product

Customer reviews and Q&As

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

Application
Western blot
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
CHO cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jun 23 2015

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (Transfected ES cells)
Specification
Transfected ES cells
Treatment
1uM all-trans retinoic acid for 24hrs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Feb 27 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 18°C
Sample
Human Cell (293T)
Specification
293T
Permeabilization
Yes - 0.5% Triton X100
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 15 2014

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