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Mouse Monoclonal 6X His tag® antibody. Suitable for IP, ELISA, Dot, WB, ICC/IF and reacts with Tag samples. Cited in 355 publications.


Images

Key facts

Isotype

IgG2b

Host species

Mouse

Storage buffer

pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPELISADotWBICC/IF
Tag
Tested
Expected
Expected
Tested
Tested

Tested
Tested

Species

Tag

Dilution info

1/200

Notes

-

Expected
Expected

Species

Tag

Dilution info

Use at an assay dependent concentration.

Notes

-

Expected
Expected

Species

Tag

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Tag

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Tag

Dilution info

1/200

Notes

-

Alternative names

Recommended products

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Mouse Monoclonal 6X His tag® antibody. Suitable for IP, ELISA, Dot, WB, ICC/IF and reacts with Tag samples. Cited in 355 publications.

Alternative names

Key facts

Isotype

IgG2b

Form

Liquid

Clonality

Monoclonal

Clone number

HIS.H8

Purification technique

Affinity purification Protein A

Specificity

Recognizes His-tagged recombinant proteins or His-tagged proteins overexpressed in cells.

ab18184 reacts to recombinant proteins containing the 6X His tag® or 10X His tag® fused to either the amino or carboxy terminus.

Concentration
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Purification notes

Purified from TCS by Protein A affinity chromatography

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

This product was changed from ascites to tissue culture supernatant on 5th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

HIS-TAG® is a trademark of EMD Biosciences, Inc.

Supplementary info

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.

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.

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.

7 product images

  • Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of a customer review submitted by Mr Chris Tracy

    Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 at 1/100 dilution.

    Lane 1 : MW Ladder (from top 120, 85, 50, 34, 25, 19 kDa)
    Lanes 2-5 : Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 from mixture of ~50 ng purified His/FLAG tagged PDCD5 and 15 μL RRL in 300 μL buffer.
    Lanes 6-9 : Purified PDCD5 from E. coli with ~60-70 ng protein.

    The lowest band in each lane is ~ 20 kDa and is PDCD5. The upper bands in lanes 2-5 are the heavy chain (~50 kDa, runs as doublet) and light chain (~25 or 26 kDa).

    Western Blot was performed with a mouse anti-FLAG primary antibody at 1/2000 dilution. An IRDye®800CW-conjugated goat anti-mouse IgG was used as a secondary antibody.

    All lanes: Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (AB18184)

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of a customer review submitted by Vladimir Milenkovic

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    Blocked with 5% milk for 30 minutes at 25°C.

    Incubated with the primary antibody for 16 hours at 4°C in 1X PBS/Tween, 5% milk.

    All lanes: Western blot - Anti-6X His tag® antibody [HIS.H8] (AB18184) at 1/5000 dilution

    Lane 1: ARPE-19 whole cell lysate - non-transfected at 20 µg

    Lane 2: ARPE-19 whole cell lysate - transfected with rRab27a-His at 20 µg

    Secondary

    All lanes: HRP-conjugated goat anti-mouse IgG polyclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 5min

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of an anonymous customer review.

    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    ab18184 at a 1/1000 dilution staining 6X His tag in transfected human HEK293 cells by Immunocytochemistry/ Immunofluorescence incubated for 12 hours at 22°C. PFA fixed. Permeabilized using 0.1% Triton X-100 in PBS. Blocked using 3% BSA for 1 hour at 22°C. Secondary used at a 1/1000 dilution polyclonal Goat anti-mouse IgG (H+L) conjugated to Alexa Fluor 488 (green). DAPI staining (blue).

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    LEFT: untransfected control; RIGHT: anti-His (in red) on His-tagged fusion proteins in HEK293 cells. Both counterstained with DAPI (in blue).

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail
    This image is courtesy of a customer review submitted by Rachel Loh

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    All lanes: Western blot - Anti-6X His tag® antibody [HIS.H8] (AB18184) at 1/1000 dilution

    Lane 1: 6x His-tagged protein expressed in BL21 (pET15b) at 1 µg

    Lane 2: 6x His-tagged protein expressed in BL21 (pET15b) at 0.1 µg

    Secondary

    All lanes: IRDye® 800CW goat anti-mouse IgG (H+L) polyclonal at 1/5000 dilution

    Performed under reducing conditions.

    Exposure time: 5min

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    ab286909 was shown to bind specifically to SARS-CoV-2 nsp13 protein in western blot. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    Blocking buffer: 5% milk in TBS-0.1% Tween® 20 (TBS-T)

    Lanes 1 - 4: Western blot - Anti-6X His tag® antibody [HIS.H8] (AB18184) at 1/1000 dilution

    Lanes 5 - 8: Western blot - Anti-SARS-CoV-2 nsp13 antibody [EPR24838-18] (AB286909) at 1/1000 dilution

    Lanes 1 and 5: Recombinant SARS-CoV-2 nsp13 protein (His-tagged) at 0.5 µg

    Lanes 2 and 6: Recombinant SARS-CoV-2 nsp13 protein (His-tagged) at 0.2 µg

    Lanes 3 and 7: Recombinant SARS-CoV2 nsp1 protein (His tagged) at 0.5 µg

    Lanes 4 and 8: Recombinant SARS-CoV2 nsp1 protein (His tagged) at 0.2 µg

    Secondary

    Lanes 1 - 4: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (AB216776) at 1/20000 dilution

    Lanes 5 - 8: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (AB216773) at 1/20000 dilution

    This data was developed using ab286909, the same antibody clone in a different buffer formulation.

    ab286909 was shown to bind specifically to SARS-CoV-2 nsp13 protein in western blot. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    Blocking buffer: 5% milk in TBS-0.1% Tween® 20 (TBS-T)

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184), expandable thumbnail

    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)

    In Western blot, ab286912 was shown to bind specifically to NSP14. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

    All lanes: Western blot - Anti-SARS-CoV-2 nsp14 antibody [EPR24839-87] (AB286912) at 1/1000 dilution

    Lane 1: Recombinant SARS-CoV-2 NSP14 protein - Active (His-Tagged) at 0.5 µg

    Lane 3: Recombinant SARS-CoV-2 NSP14 (His-tagged) at 0.5 µg

    Lane 5: SARS-CoV-2 NSP1 protein (His-tagged) at 0.5 µg

    Lane 7: Recombinant SARS-COV-2 NSP2 protein (His-tagged) at 0.5 µg

    Lane 9: Recombinant SARS-CoV-2 NSP9 Protein (His-tagged) at 0.5 µg

    Secondary

    Lanes 1 - 9: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

    Lanes 1 - 9: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 64 kDa

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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'.

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