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

Reviews (31)Q&A (10)References (169)

Key features and details

  • Mouse monoclonal [HIS.H8] to 6X His tag®
  • Suitable for: ICC/IF, ELISA, Dot blot, WB, IP
  • Reacts with: Species independent
  • Isotype: IgG2b

製品の概要

  • 製品名

    Anti-6X His tag® antibody [HIS.H8]
    6X His tag® 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [HIS.H8] to 6X His tag®

  • 由来種

    Mouse

  • 特異性

    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.

  • アプリケーション

    適用あり: ICC/IF, ELISA, Dot blot, WB, IPmore details

  • 種交差性

    交差種: Species independent

  • 免疫原

    6x His synthetic peptide.

  • ポジティブ･コントロール

  • 特記事項

    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.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    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.

  • バッファー

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • 精製度

    Protein A purified

  • 特記事項（精製）

    Purified from TCS by Protein A affinity chromatography

  • ポリ/モノ

    モノクローナル

  • クローン名

    HIS.H8

  • アイソタイプ

    IgG2b

  • 研究分野

アプリケーション

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.

アプリケーション Abreviews 特記事項
ICC/IF 1/200.
ELISA Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.
WB 1/1000.
IP 1/200.

ターゲット情報

  • 関連性

    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.

  • 細胞内局在

    Depends upon the localization of the parent protein tagged with hexahistidine.

  • 別名

    • 6 His epitope tag antibody
    • Hexa His tag antibody
    • HHHHHH epitope tag antibody
    • HHHHHH tag antibody
    • His tag antibody
    • Polyhistidine Tag antibody
    see all

画像

  • Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    Immunoprecipitation - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an Abreview submitted by Mr Chris Tracy
    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

  • Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an Abreview submitted by Vladimir Milenkovic
    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
    All lanes : HRP-conjugated goat anti-mouse IgG polyclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 5 minutes


    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

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [HIS.H8] (ab18184)
    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)
    Western blot - Anti-6X His tag® antibody [HIS.H8] (ab18184)This image is courtesy of an Abreview submitted by Rachel Loh
    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
    All lanes : IRDye® 800CW goat anti-mouse IgG (H+L) polyclonal at 1/5000 dilution

    Performed under reducing conditions.

    Exposure time: 5 minutes

    See Abreview

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

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献

ab18184 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab18184 は 169 報の論文で使用されています。

  • Li H  et al. KDM4B facilitates colorectal cancer growth and glucose metabolism by stimulating TRAF6-mediated AKT activation. J Exp Clin Cancer Res 39:12 (2020). PubMed: 31931846
  • Uzureau S  et al. APOL1 C-Terminal Variants May Trigger Kidney Disease through Interference with APOL3 Control of Actomyosin. Cell Rep 30:3821-3836.e13 (2020). PubMed: 32187552
  • Zhang X  et al. Microbiota-derived SSL6 enhances the sensitivity of hepatocellular carcinoma to sorafenib by down-regulating glycolysis. Cancer Lett 481:32-44 (2020). PubMed: 32246956
  • Bourgeois B  et al. Nonclassical nuclear localization signals mediate nuclear import of CIRBP. Proc Natl Acad Sci U S A 117:8503-8514 (2020). PubMed: 32234784
  • Alrdahe S  et al. Dysregulation of macrophage development and phenotype in diabetic human macrophages can be rescued by Hoxa3 protein transduction. PLoS One 14:e0223980 (2019). PubMed: 31626638
View all Publications for this product

レビューと Q&A

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1-10 of 41 Abreviews or Q&A

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (purified protein)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
5 µg
Treatment
fulllength, deletion mutant expression
Specification
purified protein
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 Mar 16 2020

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Fish Recombinant protein (sf9)
Gel Running Conditions
Reduced Non-Denaturing (Native) (10% SDS)
Loading amount
2e+007 cells
Specification
sf9
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Read More

Abcam user community

Verified customer

投稿 Jan 02 2020

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Escherichia coli Cell lysate - whole cell (Escherichia coli)
Gel Running Conditions
Reduced Denaturing (4-20)
Loading amount
30 µg
Specification
Escherichia coli
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Read More

Mr. Lawrence Welch

Verified customer

投稿 Jun 15 2019

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Purified protein (human, purified protein)
Gel Running Conditions
Reduced Denaturing (4-20)
Loading amount
0.25 µg
Specification
human, purified protein
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More

Abcam user community

Verified customer

投稿 Mar 20 2019

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
HEK293T
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Read More

Abcam user community

Verified customer

投稿 Jun 28 2018

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 Dec 11 2017

Immunoprecipitation abreview for Anti-6X His tag® antibody [HIS.H8]

 Good
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
10 µg
Immuno-precipitation step
Protein A/G
Specification
293T
Read More

Abcam user community

Verified customer

投稿 Oct 16 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-6X His tag® antibody [HIS.H8]

 Good
Abreviews
abreview image
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
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Abcam user community

Verified customer

投稿 Oct 13 2017

Western blot abreview for Anti-6X His tag® antibody [HIS.H8]

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 Oct 13 2017

Immunoprecipitation abreview for Anti-6X His tag® antibody [HIS.H8]

 Average
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

投稿 Mar 25 2016

1-10 of 41 Abreviews or Q&A

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