Skip to main content

Rabbit Recombinant Monoclonal 6X His tag® antibody. Carrier free. Suitable for ICC/IF, IP, ChIP, Flow Cyt, WB, IHC-P and reacts with Tag samples. Cited in 2 publications.

Be the first to review this product! Submit a review

Images

Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492), expandable thumbnail
  • ChIP - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492), expandable thumbnail
  • Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPChIPFlow CytWBIHC-P
Tag
Tested
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species

Tag

Dilution info

-

Notes

-

Tested
Tested

Species

Tag

Dilution info

-

Notes

-

Tested
Tested

Species

Tag

Dilution info

-

Notes

-

Tested
Tested

Species

Tag

Dilution info

-

Notes

-

Tested
Tested

Species

Tag

Dilution info

-

Notes

-

Tested
Tested

Species

Tag

Dilution info

-

Notes

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

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

15 products for Alternative Product

4 products for Alternative Version

Target data

Alternative names

6 His epitope tag, HHHHHH epitope tag, HHHHHH tag, Hexa His tag, Polyhistidine Tag

Recommended products

Rabbit Recombinant Monoclonal 6X His tag® antibody. Carrier free. Suitable for ICC/IF, IP, ChIP, Flow Cyt, WB, IHC-P and reacts with Tag samples. Cited in 2 publications.

Alternative names

6 His epitope tag, HHHHHH epitope tag, HHHHHH tag, Hexa His tag, Polyhistidine Tag

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR20547

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab232492 is the carrier-free version of Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204.

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

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The 6X His tag is a sequence commonly used for protein purification detection and immobilization. It consists of six histidine residues that bind to metal ions like nickel or cobalt. This makes it a highly efficient tag for affinity chromatography. His tag antibodies such as anti-His HRP facilitate the detection of His-tagged proteins in various assays. Being a small tag with a typical molecular weight of about 0.84 kDa it minimally interferes with protein function. Many proteins expressed in E. coli and other expression systems carry the 6X His tag for ease of purification and analysis.

Biological function summary

Proteins tagged with the 6X His tag often play roles in studies involving recombinant proteins. The tag itself does not alter the biological function of the target protein. It is not part of any natural protein complex but aids in the study of such complexes. Researchers use His tag antibodies like anti-His to specifically target and study these tagged proteins without cross-reactivity to other proteins.

Pathways

The incorporation of the 6X His tag into proteins allows them to be studied more efficiently in various biological pathways. It can be part of pathways like signal transduction or metabolic pathways. When tagged proteins interact with other molecules such as kinases or receptors researchers can track these interactions using the tag and anti-Histidine antibodies. This technique informs on proteins that are hard to purify due to low abundance or instability.

Associated diseases and disorders

The 6X His tag itself does not have direct implications. However proteins of interest in disease models often carry the tag which allows researchers to purify and study them in detail. For example cancer-related proteins may be expressed with a His tag to understand their role in tumorigenesis. Similarly proteins involved in metabolic disorders could be tagged which helps in analyzing specific interactions or functions disrupted in diseases.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492), expandable thumbnail

    Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    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 Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

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

    Lane 2: Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 IP in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-6X His tag® antibody [EPR20547] - ChIP Grade 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204).

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

    Observed band size: 125 kDa

  • ChIP - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492), expandable thumbnail

    ChIP - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    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 Anti-6X His tag® antibody [EPR20547] - ChIP Grade 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204).

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    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 Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204).

  • Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492), expandable thumbnail

    Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    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 Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 (right panel) at 1/5000 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    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 Anti-6X His tag® antibody [EPR20547] - ChIP Grade 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204).

    Heat mediated antigen retrieval was performed with 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] - BSA and Azide free (ab232492), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    Immunohistochemical analysis of paraffin-embedded human liver tissue labeling 6X His tag® with Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Negative control: No staining on human liver.

    Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204).

    Heat mediated antigen retrieval was performed with 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] - BSA and Azide free (ab232492), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling 6X His tag® with Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Negative control: No staining on rat stomach.

    Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    Clone EPR20547 (ab232492) has been successfully conjugated by Abcam. This image was generated using Anti-6X His tag® antibody [EPR20547] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-6X His tag® antibody [EPR20547] ab237337 for protocol details.

    Alexa Fluor® 647 Anti-6X His tag® antibody [EPR20547] ab237337 staining 6X His tag® in 293T cells transfected with 6X His tag® with GFP tag. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 647 Anti-6X His tag® antibody [EPR20547] ab237337 at 1/100 dilution (shown in red). Nuclear DNA was labeled with DAPI (shown in blue) and GFP is shown in green.

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

    Clone EPR20547 (ab232492) has been successfully conjugated by Abcam. This image was generated using Anti-6X His tag® antibody [EPR20547] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-6X His tag® antibody [EPR20547] ab237336 for protocol details.

    Alexa Fluor® 488 Anti-6X His tag® antibody [EPR20547] ab237336 staining 6X His tag® in 293T cells transfected with 6X His tag® with MYC tag. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 488 Anti-6X His tag® antibody [EPR20547] ab237336 at 1/200 dilution (shown in green) and Mouse monoclonal to Myc-Tag (Alexa Fluor® 647) (shown in red). Nuclear DNA was labeled with DAPI (shown in blue).

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com