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使用敲除细胞株进行验证重组RabMAb

重组Anti-FAK抗体[EP695Y] - BSA and Azide free (ab271836)

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Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
  • Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
  • Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
  • Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
  • Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP695Y] to FAK - BSA and Azide free
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 568 Alexa Fluor® 647 Alexa Fluor® 750 Unconjugated

概述

  • 产品名称

    Anti-FAK抗体[EP695Y] - BSA and Azide free
    参阅全部 FAK 一抗

  • 描述

    兔单克隆抗体[EP695Y] to FAK - BSA and Azide free

  • 宿主

    Rabbit

  • 特异性

    ab271836 recognises Focal adhesion kinase (FAK).

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • 经测试应用

    适用于: WB, IHC-Pmore details

  • 种属反应性

    与反应: Mouse, Rat, Human
    预测可用于: Cow

  • 免疫原

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

  • 阳性对照

    • IHC-P: human hepatocellular carcinoma

  • 常规说明

    ab271836 is the carrier-free version of ab40794.

    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ab271836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
Use at an assay dependent concentration. Predicted molecular weight: 119 kDa.

For unpurified use at 1/1000

 
IHC-P
Use at an assay dependent concentration.

The mouse, rat and cow recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and cow.

See IHC antigen retrieval protocols.
说明
WB
Use at an assay dependent concentration. Predicted molecular weight: 119 kDa.

For unpurified use at 1/1000

 
IHC-P
Use at an assay dependent concentration.

The mouse, rat and cow recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and cow.

See IHC antigen retrieval protocols.

靶标

  • 功能

    Non-receptor protein-tyrosine kinase implicated in signaling pathways involved in cell motility, proliferation and apoptosis. Activated by tyrosine-phosphorylation in response to either integrin clustering induced by cell adhesion or antibody cross-linking, or via G-protein coupled receptor (GPCR) occupancy by ligands such as bombesin or lysophosphatidic acid, or via LDL receptor occupancy. Microtubule-induced dephosphorylation at Tyr-397 is crucial for the induction of focal adhesion disassembly. Plays a potential role in oncogenic transformations resulting in increased kinase activity.

  • 组织特异性

    Expressed in all organs tested, in lymphoid cell lines, but most abundantly in brain.

  • 序列相似性

    Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
    Contains 1 FERM domain.
    Contains 1 protein kinase domain.

  • 结构域

    The first Pro-rich domain interacts with the SH3 domain of CRK-associated substrate (BCAR1) and CASL.
    The carboxy-terminal region is the site of focal adhesion targeting (FAT) sequence which mediates the localization of FAK1 to focal adhesions.

  • 翻译后修饰

    Phosphorylated on 6 tyrosine residues upon activation. Microtubule-induced dephosphorylation at Tyr-397 could be catalyzed by PTPN11 and regulated by ZFYVE21. Dephosphorylated by PTPN11 upon EPHA2 activation by its ligand EFNA1.

  • 细胞定位

    Cell junction > focal adhesion. Cell membrane. Constituent of focal adhesions.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • FADK 1 antibody
    • FADK antibody
    • FAK related non kinase polypeptide antibody
    • FAK1 antibody
    • FAK1_HUMAN antibody
    • Focal adhesion kinase 1 antibody
    • Focal adhesion Kinase antibody
    • Focal adhesion kinase isoform FAK Del33 antibody
    • Focal adhesion kinase related nonkinase antibody
    • FRNK antibody
    • p125FAK antibody
    • pp125FAK antibody
    • PPP1R71 antibody
    • Protein phosphatase 1 regulatory subunit 71 antibody
    • Protein tyrosine kinase 2 antibody
    • Protein-tyrosine kinase 2 antibody
    • Ptk2 antibody
    • PTK2 protein tyrosine kinase 2 antibody
    see all

图片

  • Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    All lanes : Anti-FAK antibody [EP695Y] (ab40794) at 1/2000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 119 kDa



    This data was developed using ab40794, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    All lanes : Anti-FAK antibody [EP695Y] (ab40794) at 1/2000 dilution

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
    Lane 2 : Rat brain lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 119 kDa



    This data was developed using ab40794, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Anti-FAK antibody [EP695Y] (ab40794) at 1/1000 dilution + Hela cell lysate

    Predicted band size: 119 kDa
    Observed band size: 119 kDa



    This data was developed using ab40794, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Western blot - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    All lanes : Anti-FAK antibody [EP695Y] (ab40794) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Wild-type HEK-293T cell lysate
    Lane 4 : PTK2 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 119 kDa



    This data was developed using ab40794, the same antibody clone in a different buffer formulation

    Lanes 1 - 4: Merged signal (red and green). Green - ab40794 observed at 119 kDa. Red - loading control, ab8245 observed at 37 kDa.

     ab40794 was shown to react with FAK in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255421 (knockout cell lysate ab263766) was used. Wild-type and FAK knockout samples were subjected to SDS-PAGE. ab40794 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular carcinoma tissue sections labeling FAK with purified ab40794 at 1:250 dilution (2.32 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody.

    Negative control: PBS instead of the primary antibody.

    Hematoxylin was used as a counterstain.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40794).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)

    Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using ab40794.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40794).

  • Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)
    Anti-FAK antibody [EP695Y] - BSA and Azide free (ab271836)

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)

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