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[IF=7.419] Li Liu. et al. Nanosilver inhibits the progression of pancreatic cancer by inducing a paraptosis-like mixed type of cell death. BIOMED PHARMACOTHER. 2022 Sep;153:113511  IHC ;  Chicken.  

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[IF=4.489] Xin-Li Xiao. et al. Status epilepticus induced Gadd45b is required for augmented dentate neurogenesis. Stem Cell Res. 2020 Dec;49:102102  IF ;  Mouse.  

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[IF=4.2] Yuan, Qing, et al. "Docetaxel-loaded solid lipid nanoparticles suppress breast cancer cells growth with reduced myelosuppression toxicity." International Journal of Nanomedicine 9 (2014): 4829.  WB ;  Mouse.  

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[IF=2.985] Qingfan Mo. et al. BTNL9 is frequently downregulated and inhibits proliferation and metastasis via the P53/CDC25C and P53/GADD45 pathways in breast cancer. Biochem Bioph Res Co. 2021 May;553:17  WB ;  Human.  

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Sample:
Lane1: Brain (Rat) Lysate at 30 ug
Lane2:Kidney (Rat) Lysate at 30 ug
Primary: Anti-GADD45 (bs-1360R) at 1:200 dilution;
Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bs-0295G-HRP) at 1: 3000 dilution;
Predicted band size : 18kD
Observed band size : 18kD
We are unsure as to the identity of these extra bands

Paraformaldehyde-fixed, paraffin embedded (Human liver carcinoma); 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 (GADD45) Polyclonal Antibody, Unconjugated (bs-1360R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue/cell: rat brain tissue; 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-GADD45 Polyclonal Antibody, Unconjugated(bs-1360R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) 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 (GADD45) Polyclonal Antibody, Unconjugated (bs-1360R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue/cell: U-2OS cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (GADD45) Polyclonal Antibody, Unconjugated (bs-1360R) 1:200, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.

Tissue/cell: A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (GADD45) Polyclonal Antibody, Unconjugated (bs-1360R) 1:200, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.

Blank control: A431.
Primary Antibody (green line): Rabbit Anti-GADD45 antibody (bs-1360R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody: Goat anti-rabbit IgG-AF647
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.