Anti-AKT2 (phospho S474)抗体(ab38513)

概述

  • 产品名称Anti-AKT2 (phospho S474)抗体
    参阅全部 AKT2 一抗
  • 描述
    兔多克隆抗体to AKT2 (phospho S474)
  • 特异性Akt2 (phospho-Ser474) antibody detects endogenous levels of Akt2 only when phosphorylated at serine 474. The region of AKT2 surrounding S474 has a high degree of similarity to the corresponding regions in AKT1 and AKT3 and thus may cross react with these proteins if phosphorylated on the corresponding serine residue.
  • 经测试应用适用于: WB, IHC-P, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human AKT2 aa 432-481 (C terminal) (phospho S474). The exact sequence is proprietary. The antiserum was produced against synthesized phosphopeptide derived from human Akt2 around the phosphorylation site of serine 474.
    Sequence:

    SEVDTRYFD DEFTAQSITI TPPDRYDSLG LLELDQRTHF PQFSYSASIR E

  • 阳性对照
    • Lung carcinoma tissue slides, A2780 cell extract and EGF treated 293 cell lysate.

性能

应用

Our Abpromise guarantee covers the use of ab38513 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 1/500 - 1/1000. Predicted molecular weight: 55 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 Use at an assay dependent concentration.

靶标

  • 功能General protein kinase capable of phosphorylating several known proteins.
  • 组织特异性Expressed in all human cell types so far analyzed.
  • 序列相似性Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 PH domain.
    Contains 1 protein kinase domain.
  • 翻译后修饰Phosphorylation on Thr-309 and Ser-474 is required for full activity.
    Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT2 ubiquitination. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Akt2 antibody
    • AKT2_HUMAN antibody
    • HIHGHH antibody
    • murine thymoma viral (v-akt) homolog-2 antibody
    • PKB antibody
    • PKB beta antibody
    • PKBB antibody
    • PKBBETA antibody
    • PRKBB antibody
    • Protein kinase Akt 2 antibody
    • Protein kinase Akt-2 antibody
    • Protein kinase B beta antibody
    • rac protein kinase beta antibody
    • RAC-BETA antibody
    • RAC-beta serine/threonine-protein kinase antibody
    • RAC-PK-beta antibody
    • v akt murine thymoma viral oncogene homolog 2 antibody
    see all

Anti-AKT2 (phospho S474) antibody 图像

  • All lanes : Anti-AKT2 (phospho S474) antibody (ab38513)

    Lane 1 : 293 cells treated for 30 min with 200ng/ml EGF
    Lane 2 : 293 cells treated for 30 min with 200ng/ml EGF with Phospho peptide


    Predicted band size : 55 kDa
  • ab38513 staining AKT2 (phospho S474) in 293T cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.1% Tween and blocked with 3% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/20 in PBS + 3% BSA) for 2 hours. An Alexa Fluor® 555-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-AKT2 (phospho S474) antibody (ab38513) at 1/500 dilution

    Lane 1 : Extract of A2780 cells.
    Lane 2 : Extract of A2780 cells + TNF alpha.
    Lane 3 : Extract of A2780 cells + UV.

    Lysates/proteins at 30 µg per lane.

    Secondary
    Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L)

    Predicted band size : 55 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)
    Lanes can be loaded with 5-30µg of total protein.
  • This image shows human lung carcinoma tissue stained with ab38513 at 1/50 dilution.

    Image on left: untreated.

    Image on right: treated with phosphopeptide at 1µg/ml (negative control).

  • ab38513 staining AKT2 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/150 in TBS + 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

Anti-AKT2 (phospho S474) antibody (ab38513)参考文献

This product has been referenced in:
  • Zhao HJ  et al. MiR-194 Deregulation Contributes To Colorectal Carcinogenesis via Targeting AKT2 Pathway. Theranostics 4:1193-208 (2014). IHC-P ; Mouse . Read more (PubMed: 25285168) »
  • Glienke W  et al. The dual PI3K/mTOR inhibitor NVP-BGT226 induces cell cycle arrest and regulates Survivin gene expression in human pancreatic cancer cell lines. Tumour Biol 33:757-65 (2012). WB ; Human . Read more (PubMed: 22170433) »

See all 8 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step FBS / BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9
Sample Human Tissue sections (Infantile fibromatosis)
Specification Infantile fibromatosis
Permeabilization No
Fixative Formaldehyde
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提交于 Jul 18 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Sample Pig Cell (PAE cells)
Specification PAE cells
Permeabilization Yes - 0.1% Tween
Fixative Formaldehyde
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提交于 May 12 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Sample Human Cell (293T)
Specification 293T
Permeabilization Yes - 0.1% Tween
Fixative Formaldehyde
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提交于 May 07 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Pig Cell lysate - whole cell (PAE cells)
Specification PAE cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Feb 25 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step FBS / BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9
Sample Mouse Tissue sections (Mouse embryo 18dpc)
Specification Mouse embryo 18dpc
Permeabilization No
Fixative Formaldehyde
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提交于 Jan 15 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse mammary cell)
Loading amount 100 µg
Specification Mouse mammary cell
Treatment 5ng/ml TGF beta for different time periods
Gel Running Conditions Reduced Denaturing (10.0% gel)
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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提交于 Aug 31 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"