GCN2 Antibody (YA417)

GCN2 Antibody (YA417) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to GCN2.

GCN2 is a metabolic-stress sensing protein kinase that phosphorylates the alpha subunit of eukaryotic translation initiation factor 2 (EIF2S1/eIF-2-alpha) under low amino acid availability, thereby activating the integrated stress response (ISR) to adapt to amino acid starvation (By similar). Phosphorylation of EIF2S1/eIF-2-alpha inhibits global cap-dependent translation while preferentially translating ISR-specific mRNAs (e.g., transcriptional activator ATF4), enabling ATF4-mediated reprogramming of amino acid biosynthetic gene expression to alleviate nutrient depletion (PubMed:32610081). GCN2 also binds uncharged tRNAs (By similar) and promotes translational induction of protein kinase PRKCH post amino acid starvation (By similar). It regulates cell cycle progression by repressing cyclin D1 mRNA translation upon unfolded protein response (UPR) activation or activating CDKN1A/p21 mRNA translation during amino acid deprivation (PubMed:26102367). In the nervous system, GCN2 contributes to synaptic plasticity consolidation, learning, and long-term memory formation (By similar), while inhibiting neurite outgrowth (By similar). It exhibits proapoptotic activity during glucose deprivation (By similar) and suppresses global protein synthesis upon UV irradiation independently of SAPK/JNK and p38 MAPK pathways (By similar). Additionally, GCN2 restricts alphavirus replication by impairing early viral mRNA translation (By similar) and modulates adaptive immunity against yellow fever virus by promoting dendritic cell autophagy and antigen presentation (PubMed:24310610).

Free

Q9P2K8

Predicted band size: 187 kDa; Observed band size: 187 kDa

Protein A affinity purified.

Cytoplasm.

Non-conjugated

Unmodified

AB_3102832

Epigenetics and Nuclear Signaling

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human

WB: 1:500-1:2000; ICC: 1:50-1:200; IHC-P: 1:50-1:200; FC: 1:50-1:100

• 
  Western blot analysis of extracts from A549 (lane 2(20μg) and A549(lane 3(40μg) using GCN2 (HY-P80138) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of Hela cells labeling GCN2 with GCN2 Antibody (HY-P80138)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GCN2 Antibody (HY-P80138) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of A549 cells labeling GCN2 with GCN2 Antibody (HY-P80138)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with GCN2 Antibody (HY-P80138) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Flow cytometric analysis of 1X10^6 MCF-7 cells labeling GCN2 Antibody(HY-P80138, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

WB, ICC/IF, IHC-P, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human GCN2.AA range:1-50.