Overview

  • Product nameAnti-6X His tag® antibody [4D11]See all 6X His tag® primary antibodies ...
  • Description
    Mouse monoclonal [4D11] to 6X His tag®
  • SpecificityThis antibody recognises all proteins which contain either an N or C-terminal HHHHHHGS epitope tag. It has been shown by a reviewer to recognise HHHHHH alone.
  • Tested applicationsFlow Cyt, Dot Blot, ELISA, WB, IP, ICC/IF more details
  • Immunogen

    Recombinant malaria protein TRAP fused at the amino-terminal end with six His tag. The coding sequence of TRAP 1.0 was amplified from P. falciparum genomic DNA (ITO4 isolate) in standard Polymerase Chain Reaction PCR (Saiki et al. 1988) using the primer combination trap 1 5'-AGTTGGATCCAGAGATGTGCAAAACA ATATAGTGG-3' and trap 2 5'-GCGAGTAAAGCTAAGCTTCCAGCTATTCCACCTGC-3', the amplified DNA sequence was cloned in the vector pDS56/RBSII,6xHis (Hochuli et al. 1988).

  • EpitopeThis antibody was shown to recognize an epitope (HHHHHHGS) encoded by the polylinker of several expression vectors (including pDS:RBSII, pQE-30, and pQE-50).
  • Positive control
  • General notesHIS-TAG® is a trademark of EMD Biosciences, Inc.

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C (add glycerol to a final volume of 50% for extra stability). Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • PurityIgG fraction
  • Clonality Monoclonal
  • Clone number4D11
  • MyelomaNS1
  • IsotypeIgG2a
  • Light chain typekappa
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab5000 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Flow Cyt Use at an assay dependent concentration.
AP Use at an assay dependent dilution.
Dot Blot Use at an assay dependent dilution.
ELISA Use at an assay dependent dilution.
WB 1/1000.
IP Use at an assay dependent concentration. See Abreview.
ICC/IF Use at an assay dependent concentration.

Target

  • RelevanceThe 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 localizationDepends 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

Anti-6X His tag® antibody [4D11] images

  • All lanes : Anti-6X His tag® antibody [4D11] (ab5000) at 1/2000 dilution

    Lane 1 : Lysates prepared from HEK-293 cells
    Lane 2 : Lysates prepared from HEK-293 cells transfected with Beta3-His (CACNB3)

    Secondary
    HRP-conjugated goat anit-mouse IgG polyclonal at 1/5000 dilution

    This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic

    See Abreview

  • ab5000 Immunoprecipitate in human HEK293 whole cell lysate. 25µg of cell lysate incubated with primary antibody (undiluted) and matrix (Protein G) for 16 hours at 4°C. For western blotting an undiluted HRP-Goat anti-rabbit IgG polyclonal was used. Beta3-His fusion protein was immunoprecipitated using anti His Ab, and specific beta3a band (56kDa) was detected using anti beta3 Ab

    Lane 1. Lysate of HEK293 cells expressing beta3-His (CACNB3)

    Lane 2. IP with anti His Ab

    Lane 3. Non bound fraction

    See Abreview

  • Anti-6X His tag® antibody [4D11] (ab5000) at 1/1000 dilution + 6X His Tagged Protein at 1 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) (ab65485) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 30 seconds
  • ab5000 staining Human HEK 293 cells transfected with CACNB3-His by ICC/IF. Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 5% serum for 20 minutes at 25°C. The primary antibody (1/250 in 1% goat serum, 0.1% TX100, 1X PBS) was incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 546 conjugated goat anti-mouse antibody (1/500) was used as the secondary.  DAPI was used for nuclei staining (blue).

    See Abreview

References for Anti-6X His tag® antibody [4D11] (ab5000)

This product has been referenced in:
  • Journo C  et al. Human T cell leukemia virus type 2 tax-mediated NF-?B activation involves a mechanism independent of Tax conjugation to ubiquitin and SUMO. J Virol 87:1123-36 (2013). ICC/IF . Read more (PubMed: 23135727) »
  • Takagi Y  et al. Human MTH3 (NUDT18) hydrolyzes the oxidized forms of guanosine and deoxyguanosine diphosphates: A comparison with MTH1 and MTH2. J Biol Chem : (2012). WB . Read more (PubMed: 22556419) »

See all 10 Publications for this product

Product Wall

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - 0.1% Triton in PBS
Sample Human Cell (H1 (HESCs) transfected with SOX2-His)
Specification H1 (HESCs) transfected with SOX2-His
Gating Strategy isotype control (black line in histogram)
Preparation Cell harvesting/tissue preparation method: accutase and accumax dissociation
Sample buffer: PBS 7.4 + 10% FBS for washing
Username

Abcam user community

Verified customer

Submitted Mar 07 2014

The main difference between ab18184 and ab5000 is the immunogen used to produce the antibodies. ab18184 was produced against a 6X His synthetic peptide, while ab5000 was raised against a recombinant protein fused to a 6X His tag. Thus, ab18184 was orig...

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Thank you for contacting us.

I can confirm that all four products, the ab5000, ab15149, ab18184 and ab81663 detect proteins, which contain either an N or C-terminal HHHHHHGS (6x) epitope tag.

Important aspects for his tag recognition ...

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Thank you for contacting us. The laboraotry sent the response below. Please let me know if you have any further questions.

I really wouldn't expect the antibody would cross-react with glycine, particularly as there is only a single glycine in ...

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Thank you for contacting us.

Please check the catalogue number ab5000; http://www.abcam.com/6X-His-tag-antibody-4D11-ab5000.html

ab97020; http://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-HL-AP-ab97020.html

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab18184

To check the status of the order please contact our Customer Se...

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Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for...

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Thank you for contacting us and your interest in our products.

The antibodies you are interested in are provided following different purification steps. The ab84041 is provides as concentrated culture supernatant, ab5000 is provides as IgG fr...

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Thank you for your reply.

As requested, a free of charge replacementwith ab18184 has been added to your next order, number *******.


I am sorry to hear the suggestions made have not improved the results on this occasion.
Unfortunately, the only lot of ab5000 we have in stock is the same as the one your customer received. I would be pleased to arrange a replacement with an a...

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