Overview

  • Product name
    Anti-HA tag antibody [4C12]
    See all HA tag primary antibodies
  • Description
    Mouse monoclonal [4C12] 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: ELISA, ICC/IF, ChIP, IP, WBmore details
  • 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.

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
    PBS pH7.2 with 50% glycerol, 1% BSA and 0.02% sodium azide
  • Concentration information loading...
  • Clonality
    Monoclonal
  • Clone number
    4C12
  • Isotype
    IgG1
  • Research areas

Applications

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
ELISA 1/1000. PubMed: 17167031
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

  • Anti-HA tag antibody [4C12] (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

  • 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.

  • 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.

     

  • 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

This product has been referenced in:
  • Peng J  et al. Drosophila Fezf coordinates laminar-specific connectivity through cell-intrinsic and cell-extrinsic mechanisms. Elife 7:N/A (2018). Read more (PubMed: 29513217) »
  • Hilmi K  et al. CTCF facilitates DNA double-strand break repair by enhancing homologous recombination repair. Sci Adv 3:e1601898 (2017). Read more (PubMed: 28560323) »
See all 23 Publications for this product

Customer reviews and Q&As

1-10 of 25 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Human osteosarcoma)
Total protein in input
1000 µg
Treatment
Plasmid transfection for 24 hrs
Immuno-precipitation step
Protein A/G
Specification
Human osteosarcoma

Miss. Can Zhou

Verified customer

Submitted Jan 28 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Myoblast)
Gel Running Conditions
Non-reduced Denaturing (10% gel)
Loading amount
40 µg
Treatment
Plasmid transfection
Specification
Myoblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

Abcam user community

Verified customer

Submitted Jan 28 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Jan 06 2016

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Mar 03 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Retro virus transduction for protein X)
Total protein in input
500 µg
Specification
Retro virus transduction for protein X
Immuno-precipitation step
Protein A/G

Abcam user community

Verified customer

Submitted Feb 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HEK 293T cells)
Total protein in input
1000 µg
Specification
HEK 293T cells
Immuno-precipitation step
Protein A

Abcam user community

Verified customer

Submitted Jan 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Loading amount
20 µg
Specification
HEK293T
Gel Running Conditions
Reduced Denaturing (12% Tris-glycine gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 17 2012

Answer

Thank you for your reply.

You can find out more about the 100% Abreview Testing Discount Program below:

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount



For all the antibodies that you are interested in, we already guarantee them to work in ChIP experiments, regardless of species. Therefore, they would not qualify for the testing discount program, but they are covered under our Abpromise.

If there is anything else I can help you with, or if you are interested in any of our other antibodies that have not been tested in ChIP, but you might be interested in them, then please let me know.

Read More

Answer

Thank you for contacting Abcam. Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More

Answer

Thank you for contacting us. Both antibodies have not yet been tested in any IHC application (neither IHC-P or IHC-Fr). We do have a testing discount program that you can use in that case. Here is some information about how the testing discount program works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. I'd be happy to issue you a discount code for one or both antibodies. Please let me know. Also, please do not hesitate to contact us if you need any more advice or information.

Read More

1-10 of 25 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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