Anti-6X His tag®抗体[4D11] (ab5000)

概述

  • 产品名称
    Anti-6X His tag®抗体[4D11]
    参阅全部 6X His tag® 一抗
  • 描述
    小鼠单克隆抗体[4D11] to 6X His tag®
  • 特异性
    This 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.
  • 经测试应用
    适用于: Dot blot, ELISA, WB, IP, ICC/IF, Other, Flow Cytmore details
  • 免疫原

    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).

  • 表位
    This antibody was shown to recognize an epitope (HHHHHHGS) encoded by the polylinker of several expression vectors (including pDS:RBSII, pQE-30, and pQE-50).
  • 阳性对照
  • 常规说明

    This antibody clone is manufactured by Abcam.

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

     

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Add glycerol to a final volume of 50% for extra stability and aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: Ascites

    Some batches contain 0.4M arginine. Please contact Scientific Support for further information.
  • Concentration information loading...
  • 纯度
    IgG fraction
  • 克隆
    单克隆
  • 克隆编号
    4D11
  • 骨髓瘤
    NS1
  • 同种型
    IgG2a
  • 轻链类型
    kappa
  • 研究领域

应用

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.

应用 Ab评论 说明
AP Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
WB 1/1000.
IP Use at an assay dependent concentration. See Abreview.
ICC/IF Use at an assay dependent concentration.
Other Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

靶标

  • 相关性
    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.
  • 细胞定位
    Depends upon the localization of the parent protein tagged with hexahistidine.
  • 别名
    • 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] 图像

  • 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

  • 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)参考文献

This product has been referenced in:
  • Zhang J  et al. DAO1 catalyzes temporal and tissue-specific oxidative inactivation of auxin in Arabidopsis thaliana. Proc Natl Acad Sci U S A 113:11010-5 (2016). WB . Read more (PubMed: 27651492) »
  • Merriman C  et al. Lipid-tuned Zinc Transport Activity of Human ZnT8 Protein Correlates with Risk for Type-2 Diabetes. J Biol Chem 291:26950-26957 (2016). Read more (PubMed: 27875315) »

See all 15 Publications for this product

Product Wall

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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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 are, the localisation of the tag (N- o...

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I really wouldn't expect the antibody would cross-react with glycine, particularly as there is only a single glycine in the epitope tag. If it recognised a single glycine it'd cross react with nearly all untagged proteins as it's a very common amino ac...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK 293 cells transfected with CACNB3-His)
Specification
HEK 293 cells transfected with CACNB3-His
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% TX100
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

提交于 Nov 20 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HEK 293 cells transfected with Beta3-His (CACNB3))
Total protein in input
25 µg
Specification
HEK 293 cells transfected with Beta3-His (CACNB3)
Immuno-precipitation step
Protein G
Username

Herr Dr. Vladimir Milenkovic

Verified customer

提交于 Nov 20 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293 cells transfected with Beta3-His (CACNB3))
Loading amount
25 µg
Specification
HEK 293 cells transfected with Beta3-His (CACNB3)
Gel Running Conditions
Reduced Denaturing (10% SDS)
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Herr Dr. Vladimir Milenkovic

Verified customer

提交于 Nov 20 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Recombinant protein (recombinant protein (human))
Loading amount
20 µg
Specification
recombinant protein (human)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

提交于 Nov 27 2006

This antibody does indeed recognise the classical 6 x His tag HHHHHH, and therefore I would fully expect it to recognise your PMAHHHHHH motif.

The immunogen of this antibody is recombinant malaria protein TRAP fused at the amino-terminal end with six his tag. It does recognize proteins which contain either an N or C-terminal HHHHHHGS epitope tag. Unfortunately, we do not know how neighbou...

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We are only able to guarentee the concentration of purified antibodies, and as this antibody is ascitic fluid it does not fall into that catagory.

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

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