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使用敲除细胞株进行验证重组RabMAb

重组Anti-Chd1抗体[EPR28727-78] - BSA and Azide free (ab318139)

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Western blot - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
  • Western blot - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
  • Immunocytochemistry/ Immunofluorescence - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
  • Flow Cytometry (Intracellular) - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
  • Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR28727-78] to Chd1 - BSA and Azide free
  • Suitable for: ICC/IF, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Unconjugated

概述

  • 产品名称

    Anti-Chd1抗体[EPR28727-78] - BSA and Azide free
    参阅全部 Chd1 一抗

  • 描述

    兔单克隆抗体[EPR28727-78] to Chd1 - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IF, WB, Flow Cyt (Intra)more details
    不适用于: ChIP-seq,IHC-P or IP

  • 种属反应性

    与反应: Human
    不与反应: Mouse, Rat

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Wild-type HEK-293T whole cell lysates, K562 whole cell lysates, His-tagged human Chd1 protein. ICC/IF: Wildtype HEK-293T cells. Flow Cyt (Intra) : Parental HEK293T(EDWT04) cells.

  • 常规说明

    ab318139 is the carrier-free version of ab318138.

    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

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab318139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/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
说明
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
应用说明
Is unsuitable for ChIP-seq,IHC-P or IP.

靶标

  • 功能

    ATP-dependent chromatin-remodeling factor which functions as substrate recognition component of the transcription regulatory histone acetylation (HAT) complex SAGA. Regulates polymerase II transcription. Also required for efficient transcription by RNA polymerase I, and more specifically the polymerase I transcription termination step. Regulates negatively DNA replication. Not only involved in transcription-related chromatin-remodeling, but also required to maintain a specific chromatin configuration across the genome. Is also associated with histone deacetylase (HDAC) activity (By similarity). Required for the bridging of SNF2, the FACT complex, the PAF complex as well as the U2 snRNP complex to H3K4me3. Functions to modulate the efficiency of pre-mRNA splicing in part through physical bridging of spliceosomal components to H3K4me3. Required for maintaining open chromatin and pluripotency in embryonic stem cells.

  • 序列相似性

    Belongs to the SNF2/RAD54 helicase family.
    Contains 2 chromo domains.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.

  • 结构域

    The 2 chromodomains are involved in the binding to the histone H3 methyllysine at position 4 (H3K4me3).

  • 细胞定位

    Nucleus. Cytoplasm. Is released into the cytoplasm when cells enter mitosis and is reincorporated into chromatin during telophase-cytokinesis.
  • Information by UniProt

  • 数据库链接

    • 别名

      • ATP dependent helicase CHD 1 antibody
      • ATP-dependent helicase CHD1 antibody
      • CHD 1 antibody
      • CHD-1 antibody
      • Chd1 antibody
      • CHD1_HUMAN antibody
      • Chromodomain helicase DNA binding protein 1 antibody
      • Chromodomain-helicase-DNA-binding protein 1 antibody
      • DKFZp686E2337 antibody
      • OTTHUMP00000222625 antibody
      see all

    图片

    • Western blot - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
      Western blot - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
      All lanes : Anti-Chd1 antibody [EPR28727-78] (ab318138) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
      Lane 2 : CHD1 knockout HEK-293T whole cell lysate
      Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast) 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

      Observed band size: 250 kDa why is the actual band size different from the predicted?


      Exposure time: 180 seconds


      This data was developed using ab318138, the same antibody clone in a different buffer formulation.

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      In Western blot, ab318138 was shown to bind specifically to Chd1. Target of interest was observed at 250 kDa in wild-type HEK-293T cell lysates (lane 1) with no signal observed at this size in CHD1 knockout cell line ab266512 (knockout cell lysate 263151) (lane 2).

      In Western blot, Anti-Vinculin antibody [EPR8185]  (ab129002) staining at 1/10000 dilution.

    • Western blot - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
      Western blot - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
      All lanes : Anti-Chd1 antibody [EPR28727-78] (ab318138) at 1/1000 dilution

      Lane 1 : His-tagged human Chd1 protein
      Lane 2 : His-tagged human Chd2 protein

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

      Observed band size: 25 kDa why is the actual band size different from the predicted?


      Exposure time: 180 seconds


      This data was developed using ab318138, the same antibody clone in a different buffer formulation.

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      This antibody does not cross-react with human Chd2.

      In Western blot, Anti-6X His tag antibody [EPR20547]-CHIP Grade  (ab213204) staining at 1/5000 dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
      Immunocytochemistry/ Immunofluorescence - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)

      This data was developed using ab318138, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CHD1 KO HEK293T (CHD1 knockout human embryonic kidney epithelial cell) ab266512 cells labelling Chd1 with ab318138 at 1/50 (10.58 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

      Confocal image showing nuclear staining in wildtype HEK-293T cells (shown in green), and negative staining in CHD1 knockout HEK-293T cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
      Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

      ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

    • Flow Cytometry (Intracellular) - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
      Flow Cytometry (Intracellular) - Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)

      This data was developed using ab318138, the same antibody clone in a different buffer formulation.

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized CHD1 konckout HEK293T (Right) / Parental HEK293T(EDWT04) (Left) cells labelling Chd1 with ab318138 at 1/500 dilution (0.1ug) /Magenta (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

      Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

    • Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)
      Anti-Chd1 antibody [EPR28727-78] - BSA and Azide free (ab318139)

    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 (0)

    发表研究结果有使用 ab318139？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab318139 尚未被引用在任何文献中。

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