For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

We use cookies to make our site as useful as possible.

Our Cookie Policy explains how you can opt-out of the cookies we use.

If you continue without changing your cookie settings, we'll assume you’re happy with this.

Continue Continue

United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Diagnostic & Therapeutic Solutions
    Custom solutions & partnerships

    Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery.

    Create custom solutions with us

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    6x-his-tag-antibody-epr20547-chip-grade-ab213204.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Tags & Cell Markers Epitope Tags His Tag
Share by email
RecombinantRabMAb

Recombinant Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question References (3)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20547] to 6X His tag® - ChIP Grade
  • Suitable for: WB, ICC/IF, IP, Flow Cyt, ChIP, IHC-P
  • Reacts with: Recombinant fragment

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

View more associated products

Overview

  • Product name

    Anti-6X His tag® antibody [EPR20547] - ChIP Grade
    See all 6X His tag® primary antibodies
  • Description

    Rabbit monoclonal [EPR20547] to 6X His tag® - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IP, Flow Cyt, ChIP, IHC-Pmore details
  • Species reactivity

    Reacts with: Recombinant fragment
  • Immunogen

    Synthetic peptide within 6X His tag®. The exact sequence is proprietary.

  • Positive control

    • WB: His-tagged mouse Cathepsin D and CDKN2A/p16INK4a proteins; HEK-293T transfected with His-tagged Staphylococcus aureus cas9, human Brachyury, human PRDM9 and human GAPDH cell lysates. IHC-P: HEK-293T cells transfected with His-tagged S.aureus cas9. ICC/IF: HEK-293T cells transfected with GFP-Myc-His. Flow Cyt: HEK-293T cells transfected with GFP-Myc-His. IP: HEK-293T cells transfected with His-tagged S.aureus cas9 cell lysate. ChIP: Chromatin from MCF7 cells transfected with 6X His-tagged GATA3.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as 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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20547
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Epitope Tags
    • His Tag
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
    • Alexa Fluor® 488 Anti-6X His tag® antibody [EPR20547] (ab237336)
    • Alexa Fluor® 647 Anti-6X His tag® antibody [EPR20547] (ab237337)
    • PE Anti-6X His tag® antibody [EPR20547] (ab237338)
    • HRP Anti-6X His tag® antibody [EPR20547] (ab237339)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

Our Abpromise guarantee covers the use of ab213204 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
WB 1/5000.
ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/5000.
ChIP Use 2 µg for 25 µg of chromatin.
IHC-P 1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

We don’t recommend this antibody for mouse in IHC. In our hands mouse tissues showed non-specific staining.

Target

  • Relevance

    The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
  • Cellular localization

    Depends upon the localization of the parent protein tagged with hexahistidine.
  • Alternative names

    • 6 His epitope tag antibody
    • Hexa His tag antibody
    • HHHHHH epitope tag antibody
    • HHHHHH tag antibody
    • His tag antibody
    • Polyhistidine Tag antibody
    see all

Images

  • Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    His-tagged Staphylococcus aureus cas9 was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125 kDa) construct, whole cell lysate with ab213204 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213204 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

    Lane 1: HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate 10 μg (Input). 

    Lane 2: ab213204 IP in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213204 in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 1 second.

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with GFP-Myc-His vector expression construct labeling 6X His tag® with ab213204 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (white). Confocal image showing positive staining on HEK-293T cells transfected with GFP-Myc-His vector expression construct.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    Immunohistochemical analysis of agarose-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. 

    Left image: Positive staining on HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct. Right image: No staining on HEK-293T transfected with an empty expression vector. 

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    Immunohistochemical analysis of paraffin-embedded human liver tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Negative control: No staining on human liver.

    Counter stained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Negative control: No staining on rat stomach.

    Counter stained with Hematoxylin.

     

  • ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    Chromatin was prepared from MCF7 (human breast adenocarcinoma cell line) cells transfected with 6X His-tagged GATA3 according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab213204 (blue), and 20µl of A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 2μg of rabbit normal IgG was added to the beads control (yellow).  The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    ChIP was performed according to the literature (PMID:22951069).

  • Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-Myc-His vector labeling 6X His tag® with ab213204 (right panel) at 1/5000 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079) at 1/2000 dilution was used as the secondary antibody.

    Gate is set between transfected and untransfected HEK-293T cells.

  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    All lanes : Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/20000 dilution

    Lane 1 : His-tagged mouse Cathepsin D recombinant protein (aa65-310; 27kDa)
    Lane 2 : His-tagged mouse CDKN2A/p16INK4a recombinant protein (aa1-168;18kDa)

    Lysates/proteins at 0.02 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Observed band size: 18,27 kDa why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    All lanes : Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/5000 dilution

    Lane 1 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with an empty expression vector, whole cell lysate
    Lane 2 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053;125kDa) construct, whole cell lysate
    Lane 3 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human Brachyury (O15178; aa1-435;50kDa) construct, whole cell lysate
    Lane 4 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human PRDM9 (Q9NQV7; aa1-894;100kDa) construct, whole cell lysate
    Lane 5 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human GAPDH (P04406; aa1- 335;36kDa) construct, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Observed band size: 100,125,36,50 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-3: 1 second; Lane 4-5: 10 seconds.

  • Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

Protocols

  • ChIP protocols
  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (3)

    Publishing research using ab213204? Please let us know so that we can cite the reference in this datasheet.

    ab213204 has been referenced in 3 publications.

    • Tai HM  et al. Dietary supplementation of recombinant antimicrobial peptide Epinephelus lanceolatus piscidin improves growth performance and immune response in Gallus gallus domesticus. PLoS One 15:e0230021 (2020). PubMed: 32160226
    • Zhang J  et al. K-RasG12V/Y40C-PI3K/AKT pathway regulates H1.4S35ph through PKA to promote the occurrence and development of osteosarcoma cancer. Artif Cells Nanomed Biotechnol 47:2048-2057 (2019). PubMed: 31126199
    • Lu X  et al. Effect and mechanism of the aß2-GP I/rhß2-GP I complex on JEG-3 cell proliferation, migration and invasion. Mol Med Rep 17:7505-7512 (2018). PubMed: 29620217

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    ELISA abreview for Anti-6X His tag® antibody [EPR20547] - ChIP Grade

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Recombinant protein (growth factor)
    Specification
    growth factor
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Type
    Direct
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 15 2018

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Get resources and offers direct to your inbox Sign up
    A-Z by research area
    • Cancer
    • Cardiovascular
    • Cell biology
    • Developmental biology
    • Epigenetics & Nuclear signaling
    • Immunology
    • Metabolism
    • Microbiology
    • Neuroscience
    • Signal transduction
    • Stem cells
    A-Z by product type
    • Primary antibodies
    • Secondary antibodies
    • Biochemicals
    • Isotype controls
    • Flow cytometry multi-color selector
    • Kits
    • Loading controls
    • Lysates
    • Peptides
    • Proteins
    • Slides
    • Tags and cell markers
    • Tools & Reagents
    Help & support
    • Support
    • Make an Inquiry
    • Protocols & troubleshooting
    • Placing an order
    • RabMAb products
    • Biochemical product FAQs
    • Training
    • Browse by Target
    Company
    • Corporate site
    • Investor relations
    • Company news
    • Careers
    • About us
    • Blog
    Events
    • Tradeshows
    • Conferences
    International websites
    • abcam.cn
    • abcam.co.jp

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    • Modern slavery statement
    © 1998-2021 Abcam plc. All rights reserved.