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    lkb1-antibody-epr19379-ab199970.pdf

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Cell Biology Cell Cycle Cell Cycle Inhibitors p53
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-LKB1 antibody [EPR19379] (ab199970)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
  • Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
  • Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
  • Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
  • Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)
  • Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)
  • Anti-LKB1 antibody [EPR19379] (ab199970)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19379] to LKB1
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Protein
Product image
Recombinant Human LKB1 protein (ab119736)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
Product image
Human STK11 (LKB1) knockout HEK-293T cell line (ab266647)

View more associated products

Overview

  • Product name

    Anti-LKB1 antibody [EPR19379]
    See all LKB1 primary antibodies
  • Description

    Rabbit monoclonal [EPR19379] to LKB1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: HEK-293T, Jurkat and K562 whole cell lysates; Human brain, human testis, fetal heart, fetal kidney and fetal testis lysates. IP: K562 and human testis lysates.
  • General notes

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19379
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • p53
    • Cancer
    • Cell cycle
    • Cell cycle inhibitors
    • p53 pathway
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • p53 pathway
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy

Associated products

  • Alternative Versions

    • Anti-LKB1 antibody [EPR19379] - BSA and Azide free (ab223146)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

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

    • Human STK11 (LKB1) knockout HEK-293T cell line (ab266647)
  • KO cell lysates

    • Human STK11 (LKB1) knockout HEK-293T cell lysate (ab257712)
  • Recombinant Protein

    • Recombinant Human LKB1 protein (ab119736)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab199970 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
1/1000. Detects a band of approximately 48, 50 kDa (predicted molecular weight: 45, 49 kDa).
IP
1/100.
Notes
WB
1/1000. Detects a band of approximately 48, 50 kDa (predicted molecular weight: 45, 49 kDa).
IP
1/100.

Target

  • Function

    Essential role in G1 cell cycle arrest. Phosphorylates and activates members of the AMPK-related subfamily of protein kinases. Tumor suppressor.
  • Tissue specificity

    Ubiquitously expressed. Strongest expression in testis and fetal liver.
  • Involvement in disease

    Defects in STK11 are a cause of Peutz-Jeghers syndrome (PJS) [MIM:175200]. PJS is a rare hereditary disease in which there is predisposition to benign and malignant tumors of many organ systems. PJS is an autosomal dominant disorder characterized by melanocytic macules of the lips, multiple gastrointestinal hamartomatous polyps and an increased risk for various neoplasms, including gastrointestinal cancer.
    Defects in STK11 have been associated with testicular tumors (TEST) [MIM:273300]. A common solid malignancy in males. Germ cell tumors of the testis constitute 95% of all testicular neoplasms.
  • Sequence similarities

    Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. LKB1 subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Phosphorylated by a cAMP-dependent protein kinase.
  • Cellular localization

    Nucleus. Cytoplasm. Relocates to the cytoplasm when bound to CAB39 and STRAD or CAB39 and ALS2CR2.
  • Target information above from: UniProt accession Q15831 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6794 Human
    • Omim: 602216 Human
    • SwissProt: Q15831 Human
    • Unigene: 515005 Human
    • Alternative names

      • hLKB1 antibody
      • Liver kinase B1 antibody
      • LKB1 antibody
      • PJS antibody
      • Polarization related protein LKB1 antibody
      • Renal carcinoma antigen NY-REN-19 antibody
      • Serine/Threonine Kinase 11 antibody
      • Serine/threonine protein kinase 11 antibody
      • Serine/threonine protein kinase LKB1 antibody
      • Serine/threonine protein kinase STK11 antibody
      • Serine/threonine-protein kinase 11 antibody
      • Serine/threonine-protein kinase LKB1 antibody
      • Serine/threonine-protein kinase XEEK1 antibody
      • Stk11 antibody
      • STK11_HUMAN antibody
      see all

    Images

    • Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
      Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
      All lanes : Anti-LKB1 antibody [EPR19379] (ab199970) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : STK11 knockout HEK-293T cell lysate
      Lane 3 : Hu testis cell lysate
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 45, 49 kDa
      Observed band size: 55 kDa why is the actual band size different from the predicted?



      Lanes 1- 4: Merged signal (red and green). Green - ab199970 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab199970 was shown to react with LKB1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266647 (knockout cell lysate ab257712) was used. Wild-type HEK-293T and STK11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab199970 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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.

    • Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
      Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg) 
      Lane 2: STK11 knockout HAP1 whole cell lysate (20 µg) 
      Lane 3: Hu Testis whole cell lysate (20 µg) 

      Lanes 1 - 3: Merged signal (red and green). Green - ab199970 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab199970 was shown to specifically react with STK11 in wild-type HAP1 cells. No band was observed when STK11 knockout samples were examined. Wild-type and STK11 knockout samples were subjected to SDS-PAGE.  Ab199970 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
      Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
      All lanes : Anti-LKB1 antibody [EPR19379] (ab199970) at 1/1000 dilution

      Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
      Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
      Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) 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

      Predicted band size: 45, 49 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Negative control: HeLa (PMID 11297520; PMID24086281; PMID 19340305).

    • Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
      Western blot - Anti-LKB1 antibody [EPR19379] (ab199970)
      All lanes : Anti-LKB1 antibody [EPR19379] (ab199970) at 1/1000 dilution

      Lane 1 : Human brain lysate
      Lane 2 : Human fetal heart lysate
      Lane 3 : Human fetal kidney lysate
      Lane 4 : Human fetal testis lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 45, 49 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

       

    • Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)
      Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)

      LKB1 was immunoprecipitated from 1 mg of K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate with ab199970 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab199970 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: K562  whole cell lysate, 10 μg (Input).

      Lane 2: ab199970 IP in K562  whole cell lysate.

      Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab199970 in K562 whole cell lysate.

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

      Exposure time: 3 minutes.

      LKB1 isoforms: PMID 18854309.

    • Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)
      Immunoprecipitation - Anti-LKB1 antibody [EPR19379] (ab199970)

      LKB1 was immunoprecipitated from 1 mg of Human testis lysate with ab199970 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab199970 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: Human testis lysate, 10 μg (Input).

      Lane 2: ab199970 IP in Human testis lysate .

      Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab199970 in Human testis lysate .

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

      Exposure time: 3 minutes.

      LKB1 isoforms: PMID 18854309.

    • Anti-LKB1 antibody [EPR19379] (ab199970)
      Anti-LKB1 antibody [EPR19379] (ab199970)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (4)

    Publishing research using ab199970? Please let us know so that we can cite the reference in this datasheet.

    ab199970 has been referenced in 4 publications.

    • Gong L  et al. Human ESC-sEVs alleviate age-related bone loss by rejuvenating senescent bone marrow-derived mesenchymal stem cells. J Extracell Vesicles 9:1800971 (2020). PubMed: 32944188
    • Li Y  et al. CD36 plays a negative role in the regulation of lipophagy in hepatocytes through an AMPK-dependent pathway. J Lipid Res N/A:N/A (2019). PubMed: 30662007
    • Zhang H  et al. Genistein protects against ox-LDL-induced senescence through enhancing SIRT1/LKB1/AMPK-mediated autophagy flux in HUVECs. Mol Cell Biochem N/A:N/A (2018). PubMed: 30443855
    • Qiu B  et al. EMT induced by loss of LKB1 promotes migration and invasion of liver cancer cells through ZEB1-induced YAP signaling. Oncol Lett 16:6465-6471 (2018). PubMed: 30405784

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