Anti-HA tag antibody - ChIP Grade (ab9110)
Rabbit polyclonal HA tag antibody. Validated in WB, IP, ELISA, ICC, Flow Cyt, ChIP, ChIP/Chip, ICC/IF. Cited in 509 publication(s). Independently reviewed in 53 review(s).
- Datasheet
- References (641)
- Protocols
Overview
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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 -
Species reactivity
Reacts with: Species independent -
Immunogen
YPYDVPDYA (influenza hemagglutinin-HA-epitope) conjugated to KLH.
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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.
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General notes
Properties
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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
Associated products
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
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. |
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| ChIP | Use 3 µg for 25 µg of chromatin. |
Target
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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
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody - ChIP Grade (ab9110)This image is courtesy of an Abreviewab9110 staining HA-tagged proteins in HeLa cells by ICC/IF (Immunocytochemistry/immunoflurescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% saponin and blocked with 3% serum for 30 minutes at 37°C. Samples were incubated with primary antibody (2 µg/ml) in 1x PBS for 1 hour at 37°C. An Alexa Fluor® 488-conjugated Goat polyclonal to rabbit was used as secondary antibody.
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Immunoprecipitation - Anti-HA tag antibody - ChIP Grade (ab9110)This image is courtesy of an anonymous Abreviewab9110 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.
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Western blot - Anti-HA tag antibody - ChIP Grade (ab9110)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 -
ChIP - Anti-HA tag antibody - ChIP Grade (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).
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Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody - ChIP Grade (ab9110)Seitz, K. et al Send to PLoS One. 2014 Jan 29;9(1):e87132. doi: 10.1371/journal.pone.0087132. eCollection 2014 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/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)
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Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody - ChIP Grade (ab9110)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. -
Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody - ChIP Grade (ab9110)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
Protocols
References
This product has been referenced in:
- Ishiguro KI et al. MEIOSIN Directs the Switch from Mitosis to Meiosis in Mammalian Germ Cells. Dev Cell N/A:N/A (2020). Read more (PubMed: 32032549) »
- Owusu M et al. Mapping the Human Kinome in Response to DNA Damage. Cell Rep 26:555-563.e6 (2019). Read more (PubMed: 30650350) »
