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重组RabMAb

重组Anti-6X His tag®抗体[EPR20547] - BSA and Azide free (ab232492)

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Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
  • ChIP - 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 - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
  • Immunocytochemistry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
  • Immunocytochemistry - 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)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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)
  • Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

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® - BSA and Azide free
  • Suitable for: ICC, ChIP, IHC-P, WB, Flow Cyt, IP
  • Reacts with: Recombinant fragment, Species independent

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 HRP PE Unconjugated

概述

  • 产品名称

    Anti-6X His tag®抗体[EPR20547] - BSA and Azide free
    参阅全部 6X His tag® 一抗

  • 描述

    兔单克隆抗体[EPR20547] to 6X His tag® - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC, ChIP, IHC-P, WB, Flow Cyt, IPmore details

  • 种属反应性

    与反应: Recombinant fragment, Species independent

  • 免疫原

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

  • 阳性对照

    • IHC-P: Agarose embedded HEK-293T cells.

  • 常规说明

    ab232492 is the carrier-free version of ab213204.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    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.

    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.

    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. Do Not Freeze.

  • 存储溶液

    pH: 7.2
    Constituent: PBS

  • 无载体

    是
  • 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ab232492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
ICC
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. 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.
WB
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
说明
ICC
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. 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.
WB
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.

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] - BSA and Azide free (ab232492)
    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).

  • ChIP - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
    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).

  • Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
    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).

  • Immunocytochemistry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
    Immunocytochemistry - 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).

  • Immunocytochemistry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
    Immunocytochemistry - 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.

  • Immunocytochemistry - Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
    Immunocytochemistry - 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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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)

    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.

  • Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)
    Anti-6X His tag® antibody [EPR20547] - BSA and Azide free (ab232492)

实验方案

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

文献 (2)

发表研究结果有使用 ab232492？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab232492 被引用在 2 文献中.

  • Heller L  et al. Secretion of proteins and antibody fragments from transiently transfected endothelial progenitor cells. J Cell Mol Med 24:8772-8778 (2020). PubMed: 32610368
  • Wang C  et al. Endocytosis of flavivirus NS1 is required for NS1-mediated endothelial hyperpermeability and is abolished by a single N-glycosylation site mutation. PLoS Pathog 15:e1007938 (2019). PubMed: 31356638

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