Anti-Phospho-GCN2 (Thr899) antibody (bs-3155R)

Anti-Phospho-GCN2 (Thr899) antibody

DATASHEET

Host:Rabbit

Target Protein:Phospho-GCN2 (Thr899)

IR:Immunogen Range:HL(p-T)GM

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:440275

Swiss Prot:Q9P2K8

Source:KLH conjugated Synthesised phosphopeptide derived from human GCN2 around the phosphorylation site of Thr899:HL(p-T)GM 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:GCN2 belongs to a family of kinases that phosphorylate the alpha subunit of eukaryotic translation initiation factor 2 to downregulate protein synthesis in response to varied cellular stresses.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
Flow-Cyt(1ug/Test)

Cross Reactive Species:Human
Mouse
Rat
Dog
Cow
Horse
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Tissue: Frozen section of mouse brain;
Incubation: Anti-Phospho-GCN2 (Thr899) Antibody, conjugated (bs-3155R-FITC);
Dilution: 1:500, 24 hours at 4°C;
The image is provided by Shanghai Institutes for Biological Sciences.

Blank control (Black line): Molt4 (Black).
Primary Antibody (green line):Rabbit Anti-Phospho-GCN2 (Thr899) antibody (bs-3155R)
Dilution:1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Blank control:Molt4.
Primary Antibody (green line): Rabbit Anti-Phospho-GCN2 (Thr899) antibody (bs-3155R-FITC)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. Acquisition of 20,000 events was performed.

Mouse macrophage cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Phospho-GCN2 (Thr899) Antibody(bs-3155R-PE)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (purple),blank(orange).Shanghai Institute of Nutrition and Health/Professor Guo’s laboratory