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

Reviews (12)Q&A (13)References (38)
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: 0.02% Thimerosal (merthiolate)
    Constituents: 50% Glycerol (glycerin, glycerine), 1% BSA, PBS
  • Concentration information loading...

  • Clonality

    Monoclonal

  • Clone number

    12CA5

  • Isotype

    IgG1

  • 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 (38)

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

ab1424 has been referenced in 38 publications.

  • Wang L  et al. Nuclear hnRNPA2B1 initiates and amplifies the innate immune response to DNA viruses. Science 365:N/A (2019). PubMed: 31320558
  • Wang Y  et al. p300 Acetylates JHDM1A to inhibit osteosarcoma carcinogenesis. Artif Cells Nanomed Biotechnol 47:2891-2899 (2019). PubMed: 31307234
  • Wang H  et al. Deletion of OSBPL2 in auditory cells increases cholesterol biosynthesis and drives reactive oxygen species production by inhibiting AMPK activity. Cell Death Dis 10:627 (2019). PubMed: 31427568
  • Xiang T  et al. Tumor suppressive BTB/POZ zinc-finger protein ZBTB28 inhibits oncogenic BCL6/ZBTB27 signaling to maintain p53 transcription in multiple carcinogenesis. Theranostics 9:8182-8195 (2019). PubMed: 31754389
  • James Theoga Raj C  et al. The copper-sensing transcription factor Mac1, the histone deacetylase Hst1, and nicotinic acid regulate de novo NAD+ biosynthesis in budding yeast. J Biol Chem 294:5562-5575 (2019). PubMed: 30760525
View all Publications for this product

Customer reviews and Q&As

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1-10 of 25 Abreviews or Q&A

Immunoprecipitation abreview for Anti-HA tag antibody [12CA5]

 Excellent
Abreviews
abreview image
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
Read More

Miss. Can Zhou

Verified customer

Submitted Jan 28 2016

Western blot abreview for Anti-HA tag antibody [12CA5]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Jan 28 2016

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

Abcam user community

Verified customer

Submitted Jan 06 2016

Western blot abreview for Anti-HA tag antibody [12CA5]

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

Abcam user community

Verified customer

Submitted Mar 03 2015

Immunoprecipitation abreview for Anti-HA tag antibody [12CA5]

 Good
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Retro virus transduction for protein X)
Total protein in input
500 µg
Immuno-precipitation step
Protein A/G
Specification
Retro virus transduction for protein X
Read More

Abcam user community

Verified customer

Submitted Feb 26 2013

Immunoprecipitation abreview for Anti-HA tag antibody [12CA5]

 Good
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HEK 293T cells)
Total protein in input
1000 µg
Immuno-precipitation step
Protein A
Specification
HEK 293T cells
Read More

Abcam user community

Verified customer

Submitted Jan 16 2013

Western blot abreview for Anti-HA tag antibody [12CA5]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing (12% Tris-glycine gel)
Loading amount
20 µg
Specification
HEK293T
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Sep 17 2012

Question

Thanks very much for your quick reply. I'm interested in the testing discount program you mentioned. Could you please let me know more about it?

Read More
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

Question

Dear Sir/Madam, I'm doing a ChIP experiment using drosophila tissue. Your ChIP-grade anti-epitope tag antibodies may be very helpful for me. The Abcam antibodies usually work well for us, so I would like to give it a try. But before we place a order, I'm wondering whether I can get a small aliquot of the antibodies to see whether they would work for my tissue. I'm particularly interested in AbID 9110 (or 1424) and 9132. Please let me know. hank you very much in advance!

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

Question

Our lab is trying to use these antibodies (ab9110 and ab1424) to do immunohistochemistry on sections from various mouse organs. We were wondering if these antibodies are tested and approved for use on paraffin sections only, cryopreserved sections only, or both?

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

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