AB232492

Anti-6X His tag® 抗体 [EPR20547] - BSA and Azide free

Anti-6X His tag® antibody [EPR20547] - BSA and Azide free

RabMAb
Recombinant
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(2 Publications)

Rabbit Recombinant Monoclonal 6X His tag® antibody. Carrier free. Suitable for ICC/IF, IP, ChIP, Flow Cyt, WB, IHC-P and reacts with Tag samples. Cited in 2 publications.

別名を表示する

6 His epitope tag, HHHHHH epitope tag, HHHHHH tag, Hexa His tag, Polyhistidine Tag, 6x his, 6x-HIS

9 Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Negative control : No staining on human liver.

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213204).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Clone EPR20547 (ab232492) has been successfully conjugated by Abcam. This image was generated using Anti-6X His tag® antibody [EPR20547] (Alexa Fluor® 647). Please refer to ab237337 for protocol details.

ab237337 staining 6X His tag^® in 293T cells transfected with 6X His tag^® with GFP tag. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab237337 at 1/100 dilution (shown in red). Nuclear DNA was labeled with DAPI (shown in blue) and GFP is shown in green.

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunohistochemical analysis of agarose-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Left image : Positive staining on HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct. Right image : No staining on HEK-293T transfected with an empty expression vector.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213204).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Flow Cyt

Supplier Data

Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-Myc-His vector labeling 6X His tag® with ab213204 (right panel) at 1/5000 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (ab150079) at 1/2000 dilution was used as the secondary antibody.

Gate is set between transfected and untransfected HEK-293T cells.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213204).

ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Clone EPR20547 (ab232492) has been successfully conjugated by Abcam. This image was generated using Anti-6X His tag® antibody [EPR20547] (Alexa Fluor® 488). Please refer to ab237336 for protocol details.

ab237336 staining 6X His tag^® in 293T cells transfected with 6X His tag^® with MYC tag. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab237336 at 1/200 dilution (shown in green) and Mouse monoclonal to Myc-Tag (Alexa Fluor^® 647) (shown in red). Nuclear DNA was labeled with DAPI (shown in blue).

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with GFP-Myc-His vector expression construct labeling 6X His tag® with ab213204 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (white). Confocal image showing positive staining on HEK-293T cells transfected with GFP-Myc-His vector expression construct.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213204).

Supplier Data

Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

His-tagged Staphylococcus aureus cas9 was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125 kDa) construct, whole cell lysate with ab213204 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213204 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate 10 μg (Input).

Lane 2 : ab213204 IP in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab213204 in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 1 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213204).

All lanes:

Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (<a href='/products/primary-antibodies/6x-his-tag-antibody-epr20547-chip-grade-ab213204'>ab213204</a>)

Observed band size: 125 kDa

false

ChIP - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

ChIP

Supplier Data

ChIP - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Chromatin was prepared from MCF7 (human breast adenocarcinoma cell line) cells transfected with 6X His-tagged GATA3 according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab213204 (blue), and 20µl of A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

ChIP was performed according to the literature (PMID : 22951069).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213204).

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Negative control : No staining on rat stomach.

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213204).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Key facts

宿主種

Rabbit

クローン性

Monoclonal

クローン番号

EPR20547

アイソタイプ

IgG

キャリアフリー

Yes

アプリケーション

WB, ChIP, IHC-P, ICC/IF, IP, Flow Cyt

applications

免疫原

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "ChIP" : {"fullname" : "ChIP", "shortname":"ChIP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Tag": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "ChIP-species-checked": "testedAndGuaranteed", "ChIP-species-dilution-info": "", "ChIP-species-notes": "", "FlowCyt-species-checked": "testedAndGuaranteed", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "We don't recommend this antibody for mouse in IHC. In our hands mouse tissues showed non-specific staining. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." } } }

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製品の詳細

ab232492 is the carrier-free version of ab213204.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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出荷温度及び保存条件

製品の状態

Liquid

精製方法

Affinity purification Protein A

バッファー組成

pH: 7.2 - 7.4 Constituents: PBS

出荷温度

Blue Ice

短期保存温度

+4°C

長期保存温度

+4°C

保管に関する情報

Do Not Freeze

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補足情報

This supplementary information is collated from multiple sources and compiled automatically.

The 6X His tag is a sequence commonly used for protein purification detection and immobilization. It consists of six histidine residues that bind to metal ions like nickel or cobalt. This makes it a highly efficient tag for affinity chromatography. His tag antibodies such as anti-His HRP facilitate the detection of His-tagged proteins in various assays. Being a small tag with a typical molecular weight of about 0.84 kDa it minimally interferes with protein function. Many proteins expressed in E. coli and other expression systems carry the 6X His tag for ease of purification and analysis.

Biological function summary

Proteins tagged with the 6X His tag often play roles in studies involving recombinant proteins. The tag itself does not alter the biological function of the target protein. It is not part of any natural protein complex but aids in the study of such complexes. Researchers use His tag antibodies like anti-His to specifically target and study these tagged proteins without cross-reactivity to other proteins.

Pathways

The incorporation of the 6X His tag into proteins allows them to be studied more efficiently in various biological pathways. It can be part of pathways like signal transduction or metabolic pathways. When tagged proteins interact with other molecules such as kinases or receptors researchers can track these interactions using the tag and anti-Histidine antibodies. This technique informs on proteins that are hard to purify due to low abundance or instability.

The 6X His tag itself does not have direct implications. However proteins of interest in disease models often carry the tag which allows researchers to purify and study them in detail. For example cancer-related proteins may be expressed with a His tag to understand their role in tumorigenesis. Similarly proteins involved in metabolic disorders could be tagged which helps in analyzing specific interactions or functions disrupted in diseases.

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製品プロトコール

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information 6X His tag®

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文献 (2)

Recent publications for all applications. Explore the full list and refine your search

Journal of cellular and molecular medicine 24:8772-8778 PubMed32610368

2020

Secretion of proteins and antibody fragments from transiently transfected endothelial progenitor cells.

Applications

Unspecified application

Species

Unspecified reactive species

Loree Heller,Reynald Thinard,Melanie Chevalier,Sezgi Arpag,Yu Jing,Ruth Greferath,Richard Heller,Claude Nicolau

PLoS pathogens 15:e1007938 PubMed31356638

2019

Endocytosis of flavivirus NS1 is required for NS1-mediated endothelial hyperpermeability and is abolished by a single N-glycosylation site mutation.

Applications

Unspecified application

Species

Unspecified reactive species

Chunling Wang,Henry Puerta-Guardo,Scott B Biering,Dustin R Glasner,Edwina B Tran,Mark Patana,Trent A Gomberg,Carmel Malvar,Nicholas T N Lo,Diego A Espinosa,Eva Harris

View all publications

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Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-6X His tag® 抗体 [EPR20547] - BSA and Azide free

Anti-6X His tag® antibody [EPR20547] - BSA and Azide free

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

ChIP - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (AB232492)

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Key facts

宿主種

クローン性

クローン番号

アイソタイプ

キャリアフリー

アプリケーション

免疫原

代替製品

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組合せ製品

Sample Prep & Detection Kits

Sample Prep & Detection Kits

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Secondary Antibodies

Reactivity data

製品の詳細

出荷温度及び保存条件

製品の状態

精製方法

バッファー組成

出荷温度

短期保存温度

長期保存温度

保管に関する情報

補足情報

Biological function summary

Pathways

製品プロトコール

ターゲットの情報

文献 (2)

Abcam product promise