使用敲除细胞株进行验证

Anti-LKB1抗体[Ley 37D/G6] (ab15095)

概述

  • 产品名称
    Anti-LKB1抗体[Ley 37D/G6]
    参阅全部 LKB1 一抗
  • 描述
    小鼠单克隆抗体[Ley 37D/G6] to LKB1
  • 特异性
    This antibody detects a single clean band representing LKB1/STK11 in Western blots on cells expressing LKB1.
  • 经测试应用
    适用于: WBmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Recombinant full length protein corresponding to LKB1.

  • 阳性对照
    • Adenocarcinoma H441 cell line, Lung carcinoma cell line H1299

性能

应用

Our Abpromise guarantee covers the use of ab15095 in the following tested applications.

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

应用 Ab评论 说明
WB 1/100. Detects a band of approximately 50 kDa (predicted molecular weight: 48.6 kDa).

靶标

  • 功能
    Essential role in G1 cell cycle arrest. Phosphorylates and activates members of the AMPK-related subfamily of protein kinases. Tumor suppressor.
  • 组织特异性
    Ubiquitously expressed. Strongest expression in testis and fetal liver.
  • 疾病相关
    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.
  • 序列相似性
    Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. LKB1 subfamily.
    Contains 1 protein kinase domain.
  • 翻译后修饰
    Phosphorylated by a cAMP-dependent protein kinase.
  • 细胞定位
    Nucleus. Cytoplasm. Relocates to the cytoplasm when bound to CAB39 and STRAD or CAB39 and ALS2CR2.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-LKB1 antibody [Ley 37D/G6] 图像



  • Predicted band size : 48.6 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: LKB1 knockout HAP1 cell lysate (20 µg)

    Lanes 1 and 2: Merged signal (red and green). Green - ab15095 observed at 50 kDa. Red - loading control, ab181602, observed at 37 kDa.
    ab15095 was shown to recognize LKB1 when LKB1 knockout samples were used, along with additional cross-reactive bands. Wild-type and LKB1 knockout samples were subjected to SDS-PAGE. ab15095 and ab181602 (loading control to LKB1) were diluted 1/100 and 1/10 000 and incubated overnight at 4°C. Blots were developed withGoat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab15095 staining LKB1 in human hepatocarcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a Citrate buffer ph 6 for 20 minutes at 97ºC. Samples were incubated with primary antibody (1/100 in diluent) for 1 hour at 37°C. A HRP-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-LKB1 antibody [Ley 37D/G6] (ab15095) at 1/1000 dilution

    Lane 1 : As above
    Lane 2 : As above

    Secondary
    Polyclonal goat anti-mouse conjugated with horse radish peroxidase.

    Predicted band size : 48.6 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)

    Image courtesy of anonymous abreviewer

    Negative control (Lane 1): LKB1 knockout MEF cells.

    Positive control (Lane 2): wildtype MEF cells.

    Blocking step: 5% milk as blocking agent for 1 hour at 25 °C.

    Incubation: 16 hours at 4 °C, using 5% milk in TBS/0.1 Tween as diluent.

     

    See Abreview

实验方案

Anti-LKB1 antibody [Ley 37D/G6] (ab15095)参考文献

This product has been referenced in:
  • Imasawa T  et al. High glucose repatterns human podocyte energy metabolism during differentiation and diabetic nephropathy. FASEB J 31:294-307 (2017). WB ; Human . Read more (PubMed: 27825100) »
  • Yao YH  et al. Attenuated LKB1-SIK1 signaling promotes epithelial-mesenchymal transition and radioresistance of non-small cell lung cancer cells. Chin J Cancer 35:50 (2016). Read more (PubMed: 27266881) »

See all 24 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (A549)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris nupage gel)
Loading amount
10 µg
Specification
A549
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Sep 12 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Hepatocarcinoma (HCC))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer ph 6, 20 min at 97ºC IN A PT LINK
Permeabilization
No
Specification
Hepatocarcinoma (HCC)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Username

Miss. Virginia Gutiérrez

Verified customer

提交于 Jan 07 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (MEF)
Specification
MEF
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Sep 10 2013

Thank you for contacting us.

I have performed a sequence alignment for LKB1 for Dictyostelium and for human (which is used to raise most of our antibodies.) Unfortunately there is only a total of 33% homology between the two proteins. I have...

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I have found the amino acid sequence for LKB1 for Dictyostelium which can be found at http://www.uniprot.org/uniprot/Q15831.fasta

I have contacted the lab to see if any of our antibodies would be compat...

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Thank you for contacting us.

I had a quick chat with my colleague in lab. They have confirmed that the antibody has been raised against the full length protein so they do not have determined the epitope this antibody is recognizing

I ...

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Thank you for contacting us.

I have checked with supplying the lab, but unfortunately, the immunogen is not available for sale.

Alternatively, I could recommend an isotype control (mouse IgG2b) to be used as negative control, for i...

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Thank you for contacting us.

The immunogen was ahighly purified recombinant full length protein made in E. coli, but is not a peptide. I will check with the lab if the immunogen might be commercially available and let you know what I find ou...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (neurons)
Loading amount
50 µg
Specification
neurons
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jan 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (neurons)
Specification
neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% triton-x-100
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Jan 31 2011

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