Name: Anti-6X His tag® antibody [HIS.H8]
SKU: ab18184
Rating: 5 (32 reviews)

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

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
- ウェスタン・ブロット用ポジティブコントロールRecombinant ENO1 protein
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

Alternative Versions
- Biotin Anti-6X His tag® antibody [HIS.H8] (ab173828)
Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
Isotype control
- Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)

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.

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

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 6-9 : 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.
See Abreview
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)

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)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)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

Immunoprecipitation protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

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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レビューを送る質問を送る

1-10 of 42 Abreviews or Q&A

Application

Western blot

Sample

Human Recombinant protein (Overexpressed his-tagged protein in a eColi cell c)

Gel Running Conditions

Reduced Denaturing

Loading amount

1 µg

Specification

Overexpressed his-tagged protein in a eColi cell c

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 35°C

Abcam user community

Verified customer

投稿 Aug 13 2020

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

Abcam user community

Verified customer

投稿 Mar 16 2020

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

Abcam user community

Verified customer

投稿 Jan 02 2020

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

Mr. Lawrence Welch

Verified customer

投稿 Jun 15 2019

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

Abcam user community

Verified customer

投稿 Mar 20 2019

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

Abcam user community

Verified customer

投稿 Jun 28 2018

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

Abcam user community

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 Oct 13 2017

1-10 of 42 Abreviews or Q&A

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Key features and details

製品の概要

製品名

製品の詳細

由来種

特異性

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

特記事項（精製）

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Isotype control

アプリケーション

ターゲット情報

関連性

細胞内局在

別名

画像

プロトコール

データシートおよび資料

参考文献 (169)

レビューと Q&A

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

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

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

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

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

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

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

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

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

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