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Phospho-GCN2 (T899) Recombinant Rabbit Monoclonal Antibody [JE00-51]

Usd: 385

Specification

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Catalog# HA722249

Phospho-GCN2 (T899) Recombinant Rabbit Monoclonal Antibody [JE00-51]

Application

  • WB

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Phospho-GCN2 (T899) Recombinant Rabbit Monoclonal Antibody [JE00-51]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phosphopeptide corresponding to residues surrounding Thr899 of Human GCN2.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB

Molecular Weight

Predicted band size: 187 kDa

Positive Control

HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, Neuro-2a treated with 100nM Calyculin A for 30 minutes cell lysate, C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate.

Conjugation

unconjugated

Clone Number

JE00-51

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

Target

Function

The family of stress-responsive protein kinases include HRI (heme-regulated inhibitor or EIF2AK1), PKR (EIF2AK2 or TIK), PERK (EIF2AK3) and GCN2 (EIF2AK4). These proteins phosphorylate the eukaryotic translation initiation factor 2α (eIF2α) on Ser 51 to regulate general and gene-specific protein synthesis. Phosphorylated eIF2α acts as an inhibitor of its guanine nucleotide exchange factor eIF2B. GCN2, a unique eIF2α kinase, exists in all eukaryotes from yeast to mammals. In mammals, expression of GCN2 is highest in liver and brain tissues. GCN2 primarily initiates the phosphorylation of eIF2α in response to UV, but has been shown to increase phosphorylation activity in response to serum starvation. Also, substitution of Asp 83 for Ala on eIF2α results in impaired phosphorylation by GCN2 and PKR, suggesting a contribution of remote residues to kinase-substrate recognition.

Background References

1. Li C et al. Amino acid catabolism regulates hematopoietic stem cell proteostasis via a GCN2-eIF2alpha axis. Cell Stem Cell. 2022 Jul

2. Missiaen R et al. GCN2 inhibition sensitizes arginine-deprived hepatocellular carcinoma cells to senolytic treatment. Cell Metab. 2022 Aug

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: Q9P2K8 Human

SwissProt: Q9QZ05 Mouse

Entrez Gene: 114859 Rat

Synonyms

E2AK4_HUMAN antibody

Eif2ak4 antibody

Eukaryotic Translation Initiation Factor 2 alpha kinase 4 antibody

Eukaryotic translation initiation factor 2-alpha kinase 4 antibody

GCN2 antibody

GCN2 eIF2alpha kinase antibody

GCN2 like protein antibody

GCN2-like protein antibody

KIAA1338 antibody

MGCN2 antibody

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-GCN2 (T899) on different lysates with Rabbit anti-Phospho-GCN2 (T899) antibody (<a href="/products/HA722249" style="font-weight: bold;text-decoration: underline;">HA722249</a>) at 1/1,000 dilution and pan GCN2 antibody (<a href="/products/ET1704-68" style="font-weight: bold;text-decoration: underline;">ET1704-68</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (25 µg/Lane)<br />Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate (25 µg/Lane)<br />Lane 3: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour (25 µg/Lane)<br /><br />Predicted band size: 187 kDa<br />Observed band size: 187 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722249" style="font-weight: bold;text-decoration: underline;">HA722249</a>) at 1/1,000 dilution and pan GCN2 antibody (<a href="/products/ET1704-68" style="font-weight: bold;text-decoration: underline;">ET1704-68</a>) at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-GCN2 (T899) on different lysates with Rabbit anti-Phospho-GCN2 (T899) antibody (HA722249) at 1/1,000 dilution and pan GCN2 antibody (ET1704-68) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (25 µg/Lane)
    Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate (25 µg/Lane)
    Lane 3: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour (25 µg/Lane)

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

    Exposure time: 3 minutes; ECL: K1802;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722249) at 1/1,000 dilution and pan GCN2 antibody (ET1704-68) at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-GCN2 (T899) on different lysates with Rabbit anti-Phospho-GCN2 (T899) antibody (<a href="/products/HA722249" style="font-weight: bold;text-decoration: underline;">HA722249</a>) at 1/2,000 dilution.<br /><br />Lane 1: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 2: Neuro-2a treated with 100nM Calyculin A for 30 minutes cell lysate (20 µg/Lane)<br />Lane 1: C6 cell lysate (20 µg/Lane)<br />Lane 2: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate (20 µg/Lane)<br /><br />Predicted band size: 187 kDa<br />Observed band size: 187 kDa<br />Exposure time: 59 seconds; ECL: K1802;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722249" style="font-weight: bold;text-decoration: underline;">HA722249</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-GCN2 (T899) on different lysates with Rabbit anti-Phospho-GCN2 (T899) antibody (HA722249) at 1/2,000 dilution.

    Lane 1: Neuro-2a cell lysate (20 µg/Lane)
    Lane 2: Neuro-2a treated with 100nM Calyculin A for 30 minutes cell lysate (20 µg/Lane)
    Lane 1: C6 cell lysate (20 µg/Lane)
    Lane 2: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate (20 µg/Lane)

    Predicted band size: 187 kDa
    Observed band size: 187 kDa
    Exposure time: 59 seconds; ECL: K1802;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722249) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

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

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Conjugate: unconjugated