Product nameAnti-GCN1 antibody
DescriptionRabbit polyclonal to GCN1
Tested applicationsSuitable for: ICC/IF, WB, IP, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Rabbit, Horse, Cow, Pig, Chimpanzee, Gorilla, Bat, Elephant
Synthetic peptide corresponding to Human GCN1.
- HeLa cytoplasmic lysate 293T cytoplasmic lysate
Protein previously labeled as Gcn1l1.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.09% Sodium azide
Constituents: 0.1% BSA, Tris buffered saline
Concentration information loading...
PurityImmunogen affinity purified
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.
|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.|
FunctionActs 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.
Tissue specificityUbiquitously expressed (PubMed:9039502). Expressed in skeletal muscules, ovary and testis (PubMed:9234705).
Sequence similaritiesBelongs to the GCN1 family.
Contains 47 HEAT repeats.
Cellular localizationCytoplasm. Associates with ribosomes in undifferentiated neuroblastoma cells and increases after neuronal differentiation.
- Information by UniProt
- 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
All lanes : Anti-GCN1 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 GCN1 by Immunoprecipitation, using ab86139 at 3µg/mg lysate. Image shows immunoprecipitated GCN1 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.
This product has been referenced in:
- Kaplan A et al. Small-Molecule Stabilization of 14-3-3 Protein-Protein Interactions Stimulates Axon Regeneration. Neuron 93:1082-1093.e5 (2017). Read more (PubMed: 28279353) »
- 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) »