Anti-6X His tag®抗体[HIS.H8] (ab18184)

概述

  • 产品名称
    Anti-6X His tag®抗体[HIS.H8]
    参阅全部 6X His tag® 一抗
  • 描述
    小鼠单克隆抗体[HIS.H8] to 6X His tag®
  • 特异性
    Recognizes His-tagged recombinant proteins or His-tagged proteins overexpressed in cells.
  • 经测试应用
    适用于: ICC/IF, ELISA, Dot blot, WB, IP, EMSAmore details
  • 免疫原

    6x His synthetic peptide.

  • 阳性对照
  • 常规说明

     

    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.

    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). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液
    Preservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.2
  • Concentration information loading...
  • 纯度
    Protein A purified
  • 纯化说明
    Purified from TCS by Protein A affinity chromatography
  • 克隆
    单克隆
  • 克隆编号
    HIS.H8
  • 同种型
    IgG2b
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab18184 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
ICC/IF 1/200.
ELISA Use at an assay dependent dilution.
Dot blot Use at an assay dependent dilution.
WB 1/1000.
IP 1/200.
EMSA Use at an assay dependent dilution. PubMed: 20606007

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

图片

  • All lanes : Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/5000 dilution

    Lane 1 : ARPE-19 whole cell lysate - non-transfected
    Lane 2 : ARPE-19 whole cell lysate - transfected with rRab27a-His

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated goat anti-mouse IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Vladimir Milenkovic

    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.

    See Abreview

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

  • 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 6-7 : 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.

    See Abreview

  • 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).

    See Abreview

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

    Performed under reducing conditions.

    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Rachel Loh

    See Abreview

文献

This product has been referenced in:
  • Dickens D  et al. Modulation of LAT1 (SLC7A5) transporter activity and stability by membrane cholesterol. Sci Rep 7:43580 (2017). WB . Read more (PubMed: 28272458) »
  • Wang BD  et al. Alternative splicing promotes tumour aggressiveness and drug resistance in African American prostate cancer. Nat Commun 8:15921 (2017). IP ; Mouse . Read more (PubMed: 28665395) »

See all 72 Publications for this product

客户评价及客户问答

Application
Western blot
Sample
Human Cell lysate - whole cell (normal fibroblasts)
Gel Running Conditions
Reduced Denaturing
Loading amount
100 µg
Treatment
MG132, and UVA
Specification
normal fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 1°C
Username

Abcam user community

Verified customer

提交于 Dec 11 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
10 µg
Immuno-precipitation step
Protein A/G
Specification
293T
Username

Abcam user community

Verified customer

提交于 Oct 16 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela cells)
Permeabilization
Yes - 0.1 % Triton X100
Specification
Hela cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Oct 13 2017

Application
Western blot
Sample
Human Recombinant protein (Recombinant protein)
Gel Running Conditions
Non-reduced Denaturing (12)
Loading amount
0.1 µg
Specification
Recombinant protein
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Oct 13 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Embryonic fibrobalst cells)
Total protein in input
1e+006 cells
Treatment
Influenza virus infection
Immuno-precipitation step
Other - Protein G - Dynabeads
Specification
Embryonic fibrobalst cells
Username

Abcam user community

Verified customer

提交于 Mar 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spinal Cord (Dorsal Horn))
Antigen retrieval step
None
Permeabilization
Yes - 1% Triton, 0.25% Tween in 1XPBS
Specification
Spinal Cord (Dorsal Horn)
Blocking step
Mouse-on-Mouse Block (Vector Labs) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Feb 11 2016

Application
Western blot
Sample
Human Purified protein (S2 cell)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
0.001 µg
Specification
S2 cell
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 27 2015

Application
Western blot
Loading amount
1 µg
Gel Running Conditions
Reduced Denaturing (12% hand cast gel)
Sample
Escherichia coli Recombinant protein (6x His-tagged protein expressed in BL21 (pET15b))
Specification
6x His-tagged protein expressed in BL21 (pET15b)
Blocking step
Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Miss. Rachel Loh

Verified customer

提交于 Jan 15 2015

Application
IHC - Wholemount
Sample
Xenopus tropicalis Embryo (whole embryo)
Specification
whole embryo
Username

Abcam user community

Verified customer

提交于 Aug 04 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
10 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 01 2013

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

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