Anti-AKT2 (phospho S474)抗体(ab110231)

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: Eli-pair, WB, IHC-Pmore details
  • 种属反应性
    与反应: Human
    预测可用于: Mouse, Rat, Horse, Chicken, Dog, Pig, Chimpanzee, Gorilla, Chinese Hamster, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Human AKT2, phosphorylated at S474.

  • 阳性对照
    • This antibody gave a positive signal in LPS-treated HL60 whole cell lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human pancreas adenocarcinoma.

性能

应用

Our Abpromise guarantee covers the use of ab110231 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
Eli-pair Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). Abcam recommends using milk as the blocking agent - 3%
IHC-P Use a concentration of 5 µg/ml.

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 (ab110231) at 1 µg/ml

    Lane 1 : LPS-Treated HL60 Whole Cell Lysate
    Lane 2 : LPS-Treated HL60 Whole Cell Lysate with Immunising peptide at 1 µg/ml
    Lane 3 : LPS-Treated HL60 Whole Cell Lysate with Control peptide at 1 µg/ml

    Lysates/proteins at 25 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 55 kDa


    Exposure time : 20 minutes
    Abcam recommends using milk as the blocking agent (3%). Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab110231 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
  • IHC image of AKT2 (phospho S474) staining in Human pancreas adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110231, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab110231 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.

Anti-AKT2 (phospho S474) antibody (ab110231)参考文献

ab110231 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (macrophage)
Specification macrophage
Treatment 0.1 ug/ml LPS 15 min
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Dec 12 2014

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