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Neuroscience Neurotransmission Intracellular Signaling Kinases
使用敲除细胞株进行验证重组RabMAb

重组Anti-CrkL抗体[Y243] (ab32126)

  • Datasheet
  • SDS
Submit a review Q&A (1)References (5)

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Western blot - Anti-CrkL antibody [Y243] (ab32126)
  • Immunocytochemistry/ Immunofluorescence - Anti-CrkL antibody [Y243] (ab32126)
  • Western blot - Anti-CrkL antibody [Y243] (ab32126)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CrkL antibody [Y243] (ab32126)
  • Anti-CrkL antibody [Y243] (ab32126)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y243] to CrkL
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Human CRKL knockout HeLa cell line (ab265993)

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概述

  • 产品名称

    Anti-CrkL抗体[Y243]
    参阅全部 CrkL 一抗
  • 描述

    兔单克隆抗体[Y243] to CrkL
  • 宿主

    Rabbit
  • 经测试应用

    适用于: WB, IHC-P, ICC/IFmore details
    不适用于: Flow Cyt or IP
  • 种属反应性

    与反应: Human
  • 免疫原

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

  • 阳性对照

    • HeLa and K562 cell lysates, and human breast carcinoma
  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    Y243
  • 同种型

    IgG
  • 研究领域

    • Neuroscience
    • Neurotransmission
    • Intracellular Signaling
    • Kinases
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • Leucine Zipper
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    • Neural Signal Transduction
    • Signal Transduction
    • Adapters
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    • Cancer
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相关产品

  • Alternative Versions

    • Anti-CrkL antibody [Y243] - BSA and Azide free (ab247234)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CRKL knockout HeLa cell line (ab265993)
  • KO cell lysates

    • Human CRKL knockout HeLa cell lysate (ab257397)
  • Recombinant Protein

    • Recombinant Human CrkL protein (ab116208)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab32126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
1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 33 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50 - 1/100.
说明
WB
1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 33 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50 - 1/100.
应用说明
Is unsuitable for Flow Cyt or IP.

靶标

  • 功能

    May mediate the transduction of intracellular signals.
  • 序列相似性

    Belongs to the CRK family.
    Contains 1 SH2 domain.
    Contains 2 SH3 domains.
  • Target information above from: UniProt accession P46109 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 1399 Human
    • Omim: 602007 Human
    • SwissProt: P46109 Human
    • Unigene: 5613 Human
    • 别名

      • Crk L antibody
      • Crk like protein antibody
      • Crk-like protein antibody
      • Crkl antibody
      • CRKL_HUMAN antibody
      • Crkol antibody
      • HGNC:2363 antibody
      • Oncogene CrkL antibody
      • V crk avian sarcoma virus CT10 oncogene homolog like antibody
      • v crk sarcoma virus CT10 oncogene homolog (avian) like antibody
      • V crk sarcoma virus CT10 oncogene homolog avian like antibody
      see all

    图片

    • Western blot - Anti-CrkL antibody [Y243] (ab32126)
      Western blot - Anti-CrkL antibody [Y243] (ab32126)
      All lanes : Anti-CrkL antibody [Y243] (ab32126) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : CRKL knockout HeLa cell lysate
      Lane 3 : K-562 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 33 kDa
      Observed band size: 37 kDa why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab32126 observed at 37 kDa. Red - loading control ab7291 observed at 50 kDa.

      ab32126 Anti-CrkL antibody [Y243] was shown to specifically react with CrkL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265993 (knockout cell lysate ab257397) was used. Wild-type and CrkL knockout samples were subjected to SDS-PAGE. ab32126 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-CrkL antibody [Y243] (ab32126)
      Immunocytochemistry/ Immunofluorescence - Anti-CrkL antibody [Y243] (ab32126)
      Immunocytochemistry analysis of K-562 (human chronic myelogenous leukemia lymphoblast) labeling CrkL with purified ab32126 at 1/100 dilution (10 µg/ml). Cells were fixed with 100% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.6 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
      Negative control: PBS instead of the primary antibody.
    • Western blot - Anti-CrkL antibody [Y243] (ab32126)
      Western blot - Anti-CrkL antibody [Y243] (ab32126)
      Anti-CrkL antibody [Y243] (ab32126) at 1/1000 dilution + K562 cell lysate

      Predicted band size: 33 kDa
      Observed band size: 39 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CrkL antibody [Y243] (ab32126)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CrkL antibody [Y243] (ab32126)

      ab32126, staining human breast carcinoma by immunohistochemistry, Paraffin embedded tissue

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-CrkL antibody [Y243] (ab32126)
      Anti-CrkL antibody [Y243] (ab32126)

    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

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (5)

    发表研究结果有使用 ab32126?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab32126 被引用在 5 文献中.

    • Shi X  et al. Downregulated miR-29a promotes B cell overactivation by upregulating Crk-like protein in systemic lupus erythematosus. Mol Med Rep 22:841-849 (2020). PubMed: 32467986
    • Wang L  et al. Expression of signaling adaptor proteins predicts poor prognosis in pancreatic ductal adenocarcinoma. Diagn Pathol 12:42 (2017). PubMed: 28558797
    • Poulsen ET  et al. New Insights to Clathrin and Adaptor Protein 2 for the Design and Development of Therapeutic Strategies. Int J Mol Sci 16:29446-53 (2015). IP . PubMed: 26690411
    • Wang J  et al. SCG10 promotes non-amyloidogenic processing of amyloid precursor protein by facilitating its trafficking to the cell surface. Hum Mol Genet N/A:N/A (2013). PubMed: 23863461
    • Natsume H  et al. The CRKL gene encoding an adaptor protein is amplified, overexpressed, and a possible therapeutic target in gastric cancer. J Transl Med 10:97 (2012). WB, IHC-P ; Human . PubMed: 22591714

    客户评价及客户问答

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    Question

    I'm looking for an anti-CrkL antibody that will bind the first SH3 doman, so N-terminal of a.a. 183 (region a.a. 123-183). What epitope will these antibodies recognize?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 07 2011

    Answer

    Thank you for your inquiry. I have confirmed that neither of these two anti-CrkL antibodies, ab32018 or ab32126, bind within the region a.a. 123 - 183 (the first SH3 domain). I apologize but it doesn't look like any of the anti-CrkL antibodies that we currently carry target the first SH3 domain. You may want to try searching www.biocompare.com to find one that does. Please feel free to contact me if you have other questions.

    Read More

    Abcam Scientific Support

    回复于 Oct 07 2011

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

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