Anti-HA tag antibody [12CA5] (ab1424)

Reviews (12)Q&A (13)References (45)
Western blot - Anti-HA tag antibody [12CA5] (ab1424)
  • Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody [12CA5] (ab1424)
  • Immunoprecipitation - Anti-HA tag antibody [12CA5] (ab1424)
  • ChIP - Anti-HA tag antibody [12CA5] (ab1424)

Key features and details

  • Mouse monoclonal [12CA5] to HA tag
  • Suitable for: ICC/IF, ChIP, IP, WB
  • Reacts with: Species independent
  • Isotype: IgG1

Overview

  • Product name

    Anti-HA tag antibody [12CA5]
    See all HA tag primary antibodies

  • Description

    Mouse monoclonal [12CA5] to HA tag

  • Host species

    Mouse

  • Specificity

    The antibody detects HA-tag fused to the C-terminus or N-terminus of targeted proteins in transfected cells. The antibody works in ChIP but less well than ab9110 (see ChIP data).

  • Tested applications

    Suitable for: ICC/IF, ChIP, IP, WBmore details

  • Species reactivity

    Reacts with: Species independent

  • Immunogen

    Synthetic peptide. Derived from the human influenza hemaglutinin protein
    Database link: ADG21028

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • WB: Rab6-HA transfected HEK-293T whole cell lysate. IP: Rab6-HA transfected HEK-293T whole cell lysate. ICC/IF: Baby hamster kidney cell expressing HA-tag CFTR protein. ChIP: Xenopus laevis oocytes injected with mRNA for HA-tagged human BORIS.

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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: 0.03% Proclin 300
    Constituents: 30% Glycerol (glycerin, glycerine), 0.05% BSA, PBS, 0.019% Ethylenediaminetetraacetic acid, disodium, dihydrate
  • Concentration information loading...

  • Clonality

    Monoclonal

  • Clone number

    12CA5

  • Isotype

    IgG1

  • Light chain type

    kappa

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab1424 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
ICC/IF (1)
Use a concentration of 10 µg/ml.
ChIP
Use a concentration of 3 µg/ml. Use 3 µg per 25 µg chromatin (see figure legend).
IP (5)
Use a concentration of 10 µg/ml.
WB (6)
Use a concentration of 1 µg/ml.
Notes
ICC/IF
Use a concentration of 10 µg/ml.
ChIP
Use a concentration of 3 µg/ml. Use 3 µg per 25 µg chromatin (see figure legend).
IP
Use a concentration of 10 µg/ml.
WB
Use a concentration of 1 µg/ml.

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

  • Western blot - Anti-HA tag antibody [12CA5] (ab1424)
    Western blot - Anti-HA tag antibody [12CA5] (ab1424)This image is courtesy of an anonymous Abreview
    Anti-HA tag antibody [12CA5] (ab1424) at 1/1000 dilution + HEK293T whole cell lysate over-expressing HA-tagged Rab6 at 20 µg

    Secondary
    HRP-conjugated goat anti-mouse polyclonal IgG at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 5 seconds


    Blocked with 5% milk for 30 min at 25°C

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody [12CA5] (ab1424)
    Immunocytochemistry/ Immunofluorescence - Anti-HA tag antibody [12CA5] (ab1424)This image was kindly supplied as part of the review submitted by one of our customers.

    This picture shows baby hamster kidney cell expressing HA-tag CFTR protein. This type of cells were grown on coverslip and immunofluorescence was performed. Dilution of 1:200 was used and incubated for 2hrs with antibody.

  • Immunoprecipitation - Anti-HA tag antibody [12CA5] (ab1424)
    Immunoprecipitation - Anti-HA tag antibody [12CA5] (ab1424)Data courtesy of an anonymous AbReview

    Recombinant HA-tagged Rab6 was immunoprecipitated from 1 mg transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract with 2 μL ab1424. 

    Lane 1: Rab6-HA transfected HEK-293T whole cell extract (Input)

    Lane 2: ab1424 IP in Rab6-HA transfected HEK-293T whole cell extract.

     

  • ChIP - Anti-HA tag antibody [12CA5] (ab1424)
    ChIP - Anti-HA tag antibody [12CA5] (ab1424)

    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, 3µg of ab1424 (anti-HA, light blue)  and 3µg of ab18337 (anti-BORIS, dark blue), and 20µl of Protein A/G sepharose beads. A non-specific antibody was used as a control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).

References (45)

Publishing research using ab1424? Please let us know so that we can cite the reference in this datasheet.

ab1424 has been referenced in 45 publications.

  • Melendez J  et al. TGFß signalling acts as a molecular brake of myoblast fusion. Nat Commun 12:749 (2021). PubMed: 33531476
  • Grigsby SM  et al. Elucidating the Importance of DOT1L Recruitment in MLL-AF9 Leukemia and Hematopoiesis. Cancers (Basel) 13:N/A (2021). PubMed: 33562706
  • Lei G  et al. TRIM29 Reverses Oxaliplatin Resistance of P53 Mutant Colon Cancer Cell. Can J Gastroenterol Hepatol 2021:8870907 (2021). PubMed: 33824865
  • Shorstova T  et al. Reprogramming of Nucleotide Metabolism Mediates Synergy between Epigenetic Therapy and MAP Kinase Inhibition. Mol Cancer Ther 20:64-75 (2021). PubMed: 33087508
  • Ma L  et al. The m6A reader YTHDC2 inhibits lung adenocarcinoma tumorigenesis by suppressing SLC7A11-dependent antioxidant function. Redox Biol 38:101801 (2020). PubMed: 33232910
View all Publications for this product

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-HA tag antibody [12CA5]

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (vascular smooth muscle cell)
Permeabilization
Yes - NP40
Specification
vascular smooth muscle cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde
Read More

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Verified customer

Submitted Jan 06 2016

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