[IF=25.269] Changzheng Li. et al. Amino acid catabolism regulates hematopoietic stem cell proteostasis via a GCN2-eIF2α axis. CELL STEM CELL. 2022 Jul;29:1119  WB ;  Mouse.  

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[IF=5.58] Lehman, Stacey L., Sandra Ryeom, and Constantinos Koumenis. "Signaling through alternative Integrated Stress Response pathways compensates for GCN2 loss in a mouse model of soft tissue sarcoma." Scientific Reports 5 (2015).  WB ;  Mouse.  

PubMed:26123823

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:Mouse spleen.
Primary Antibody (green line): Rabbit Anti-Phospho-GCN2 (Thr899) antibody (bs-3155R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST 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.