[IF=4.556] Hae-Young Kim. et al. Low-Temperature Argon Plasma Regulates Skin Moisturizing and Melanogenesis-Regulating Markers through Yes-Associated Protein. Int J Mol Sci. 2021 Jan;22(4):1895  WB ;  Mouse.  

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[IF=3.234]  Kim HY et al. Inhibitory effects of extracellular superoxide dismutase on ultraviolet B-induced melanogenesis in murine skin and melanocytes. Life Sci. 2018 Oct 1;210:201-208.  WB ;  Mouse.  

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Sample:
MOLT-4(Human) Cell Lysate at 30 ug
Primary: Anti-MC1 Receptor  (bs-1419R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 35 kD

Sample:
Hela(Human) Cell Lysate at 30 ug
Raji(Human) Cell Lysate at 30 ug
Primary: Anti-MC1 Receptor  (bs-1419R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 35 kD

Tissue/cell: human melanoma; 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-MSHR Polyclonal Antibody, Unconjugated(bs-1419R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: human melanoma; 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-MSHR Polyclonal Antibody, Unconjugated(bs-1419R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control: 293T(blue).
Primary Antibody: Rabbit Anti- MC1 Receptor/AF488 Conjugated antibody (bs-1419R-AF488), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG/AF488(orange) ,used under the same conditions.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice. The cells were washed twice with 1 X PBS. The cells were incubated in 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions followed by the incubated with antibody (bs-1419R-AF488, 1μg /1x10^6 cells) for 30 min on ice. Acquisition of 20,000 events was performed.