Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
重组RabMAb

重组Anti-6X His tag®抗体[EPR20547] - ChIP Grade (ab213204)

Reviews (1) Submit a question References (19)
Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
  • Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20547] to 6X His tag® - ChIP Grade
  • Suitable for: WB, ICC/IF, IP, Flow Cyt, ChIP, IHC-P
  • Reacts with: Recombinant fragment

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Biotin Carrier Free FITC HRP PE

概述

  • 产品名称

    Anti-6X His tag®抗体[EPR20547] - ChIP Grade
    参阅全部 6X His tag® 一抗

  • 描述

    兔单克隆抗体[EPR20547] to 6X His tag® - ChIP Grade

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, ICC/IF, IP, Flow Cyt, ChIP, IHC-Pmore details

  • 种属反应性

    与反应: Recombinant fragment

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: His-tagged mouse Cathepsin D and CDKN2A/p16INK4a proteins; HEK-293T transfected with His-tagged Staphylococcus aureus cas9, human Brachyury, human PRDM9 and human GAPDH cell lysates. IHC-P: HEK-293T cells transfected with His-tagged S.aureus cas9. ICC/IF: HEK-293T cells transfected with GFP-Myc-His. Flow Cyt: HEK-293T cells transfected with GFP-Myc-His. IP: HEK-293T cells transfected with His-tagged S.aureus cas9 cell lysate. ChIP: Chromatin from MCF7 cells transfected with 6X His-tagged GATA3.

  • 常规说明

    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 production
    For more information see here.

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

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 long term. Avoid freeze / thaw cycle.

  • 存储溶液

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...

  • 纯度

    Protein A purified

  • 克隆

    单克隆

  • 克隆编号

    EPR20547

  • 同种型

    IgG

  • 研究领域

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab213204于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
1/5000.
ICC/IF
1/100.
IP
1/30.
Flow Cyt
1/5000.
ChIP
Use 2 µg for 25 µg of chromatin.
IHC-P
1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

We don't recommend this antibody for mouse in IHC. In our hands mouse tissues showed non-specific staining.
说明
WB
1/5000.
ICC/IF
1/100.
IP
1/30.
Flow Cyt
1/5000.
ChIP
Use 2 µg for 25 µg of chromatin.
IHC-P
1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

We don't recommend this antibody for mouse in IHC. In our hands mouse tissues showed non-specific staining.

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. His-tags can be fused to the amino- or carboxy- termini of 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

  • Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    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 1/10,000 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    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.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

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

    Negative control: No staining on human liver.

    Counter stained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

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

    Negative control: No staining on rat stomach.

    Counter stained with Hematoxylin.

     

  • ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    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).

  • Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

    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.

  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    All lanes : Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/20000 dilution

    Lane 1 : His-tagged mouse Cathepsin D recombinant protein (aa65-310; 27kDa)
    Lane 2 : His-tagged mouse CDKN2A/p16INK4a recombinant protein (aa1-168;18kDa)

    Lysates/proteins at 0.02 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Observed band size: 18,27 kDa why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    All lanes : Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/5000 dilution

    Lane 1 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with an empty expression vector, whole cell lysate
    Lane 2 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053;125kDa) construct, whole cell lysate
    Lane 3 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human Brachyury (O15178; aa1-435;50kDa) construct, whole cell lysate
    Lane 4 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human PRDM9 (Q9NQV7; aa1-894;100kDa) construct, whole cell lysate
    Lane 5 : HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human GAPDH (P04406; aa1- 335;36kDa) construct, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Observed band size: 100,125,36,50 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-3: 1 second; Lane 4-5: 10 seconds.

  • Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)
    Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

文献 (19)

发表研究结果有使用 ab213204？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab213204 被引用在 19 文献中.

  • Lee H  et al. Designer molecules of the synaptic organizer MDGA1 reveal 3D conformational control of biological function. J Biol Chem 299:104586 (2023). PubMed: 36889589
  • Xu Q  et al. ACL and HAT1 form a nuclear module to acetylate histone H4K5 and promote cell proliferation. Nat Commun 14:3265 (2023). PubMed: 37277331
  • Yang W  et al. A novel CTLA-4 blocking strategy based on nanobody enhances the activity of dendritic cell vaccine-stimulated antitumor cytotoxic T lymphocytes. Cell Death Dis 14:406 (2023). PubMed: 37419930
  • Reck J  et al. Small disulfide loops in peptide hormones mediate self-aggregation and secretory granule sorting. Life Sci Alliance 5:N/A (2022). PubMed: 35086936
  • Maeda R  et al. A panel of nanobodies recognizing conserved hidden clefts of all SARS-CoV-2 spike variants including Omicron. Commun Biol 5:669 (2022). PubMed: 35794202
View all Publications for this product

客户评价及客户问答

Show All 评价 Q&A
提交评价 提交问题

ELISA abreview for Anti-6X His tag® antibody [EPR20547] - ChIP Grade

 Poor
Abreviews
abreview image
Application
ELISA
Sample
Human Recombinant protein (growth factor)
Specification
growth factor
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Type
Direct
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Mar 15 2018

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com