製品の概要

  • 製品名
    Anti-Gcn1l1 antibody
  • 製品の詳細
    Rabbit polyclonal to Gcn1l1
  • アプリケーション
    適用あり: ICC/IF, WB, IP, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Rabbit, Horse, Cow, Pig, Chimpanzee, Gorilla, Bat, Elephant
  • 免疫原

    A region between residue 2621 and 2671 of human Gcn1l1 using the numbering given in entry NP_006827.1.

  • ポジティブ・コントロール
    • HeLa cytoplasmic lysate 293T cytoplasmic lysate

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • バッファー
    Preservative: 0.09% Sodium Azide
    Constituents: 0.1% BSA, Tris buffered saline
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab86139 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 µg/ml.
WB 1/2000 - 1/10000. Predicted molecular weight: 293 kDa.
IP Use at 2-5 µg/mg of lysate.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能
    Acts as a positive activator of the EIF2AK4/GCN2 protein kinase activity in response to amino acid starvation. Forms a complex with EIF2AK4/GCN2 on translating ribosomes; during this process, GCN1 seems to act as a chaperone to facilitate delivery of uncharged tRNAs that enter the A site of ribosomes to the tRNA-binding domain of EIF2AK4/GCN2, and hence stimulating EIF2AK4/GCN2 kinase activity. Participates in the repression of global protein synthesis and in gene-specific mRNA translation activation, such as the transcriptional activator ATF4, by promoting the EIF2AK4/GCN2-mediated phosphorylation of eukaryotic translation initiation factor 2 (eIF-2-alpha/EIF2S1) on 'Ser-52', and hence allowing ATF4-mediated reprogramming of amino acid biosynthetic gene expression to alleviate nutrient depletion.
  • 組織特異性
    Ubiquitously expressed (PubMed:9039502). Expressed in skeletal muscules, ovary and testis (PubMed:9234705).
  • 配列類似性
    Belongs to the GCN1 family.
    Contains 47 HEAT repeats.
  • 細胞内局在
    Cytoplasm. Associates with ribosomes in undifferentiated neuroblastoma cells and increases after neuronal differentiation.
  • Information by UniProt
  • 参照データベース
  • 別名
    • eIF-2-alpha kinase activator GCN1 antibody
    • GCN1 (general control of amino acid synthesis yeast homolog) like antibody
    • GCN1 (general control of amino acid synthesis 1 yeast) like 1 antibody
    • GCN1 antibody
    • GCN1 eIF-2-alpha kinase activator homolog antibody
    • GCN1 general control of amino acid synthesis 1 like 1 antibody
    • GCN1 general control of amino acid synthesis 1 like 1 (yeast) antibody
    • GCN1 like protein 1 antibody
    • GCN1-like protein 1 antibody
    • GCN1_HUMAN antibody
    • GCN1L antibody
    • General control of amino-acid synthesis 1-like protein 1 antibody
    • HsGCN1 antibody
    • Translational activator gcn1 antibody
    see all

画像

  • All lanes : Anti-Gcn1l1 antibody (ab86139) at 0.04 µg/ml

    Lane 1 : HeLa whole cell lysate at 50 µg
    Lane 2 : HeLa whole cell lysate at 15 µg
    Lane 3 : HeLa whole cell lysate at 5 µg
    Lane 4 : 293T whole cell lysate at 50 µg

    Developed using the ECL technique

    Predicted band size : 293 kDa
    Observed band size : 293 kDa


    Exposure time : 30 seconds
  • Detection of Human Gcn1l1 by Immunoprecipitation, using ab86139 at 3µg/mg lysate. Image shows immunoprecipitated Gcn1l1 detected with post IP WB, loading 20% of IP and using HeLa whole cell lysate at 1mg, with ab86139 at 1 µg/ml in lane 1 and control IgG in lane 2. Detection: Chemiluminescence with an exposure time of 30 seconds.
  • ICC/IF image of ab86139 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86139, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of ab86139 staining in human breast carcinoma 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 ab86139, 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.

参考文献

This product has been referenced in:
  • Vartanian S  et al. Application of Mass Spectrometry Profiling to Establish Brusatol as an Inhibitor of Global Protein Synthesis. Mol Cell Proteomics 15:1220-31 (2016). Read more (PubMed: 26711467) »

See 1 Publication for this product

レビューと Q&A

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Sample
Human Cell (293T)
Specification
293T
Permeabilization
Yes - 0.5% Triton X100
Fixative
Paraformaldehyde
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Verified customer

投稿 Sep 19 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-12 Bis tris)
Sample
Human Cell lysate - whole cell (293T)
Specification
293T
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 Sep 19 2014

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

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