[IF=3.12] Jiang, H-Q., et al. "Guanabenz Delays the Onset of Disease Symptoms, Extends Lifespan, Improves Motor Performance and Attenuates Motor Neuron Loss in the SOD1 G93A Mouse Model of Amyotrophic Lateral Sclerosis." Neuroscience (2014).  WB ;  Mouse.  

PubMed:24699224

[IF=4.86] Jiang, Mingfang, et al. "Down-regulation of miR-384-5p Attenuates Rotenone-induced Neurotoxicity in Dopaminergic SH-SY5Y Cells Through Inhibiting Endoplasmic Reticulum Stress." American Journal of Physiology-Cell Physiology (2016): ajpcell-00226.  WB ;  Human.  

PubMed:26864693

[IF=4.15] Hayashi, Keitaro, et al. "HOXB9 acts as a negative regulator of activated human T cells in response to amino acid deficiency." Immunology and Cell Biology (2016).  WB ;  Human.  

PubMed:26926958

Sample:
Embryo(Mouse) lysate at 30ug;
Eye(Mouse) lysate at 30 ug;
Primary: Anti-ATF4 (bs-1531R) at 1:300 dilution;
Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1: 5000 dilution;
Predicted band size : 38kD
Observed band size : 38kD

Paraformaldehyde-fixed, paraffin embedded (Human breast cancer); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human brain glioma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (ATF4) Polyclonal Antibody, Unconjugated (bs-1531R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.

Tissue/cell: human rectal carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-ATF4 Polyclonal Antibody, Unconjugated(bs-1531R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: human gastric carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-ATF4/CREB-2 Polyclonal Antibody, Unconjugated(bs-1531R) 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control (blue line): Hela
Primary Antibody (green line): Rabbit Anti-Bid antibody (bs-1531R)
Dilution: 0.2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.