Sample:lung(mouse) Lysate at 40 ug
Primary: Anti- CD130 (bs-1459R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 99kD
Observed band size: 130 kD

Sample: Lung(Mouse) Lysate at 30 ug
Primary: Anti- CD130 (bs-1459R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/10000 dilution
Predicted band size: 99 kD
Observed band size: 130 kD

Paraformaldehyde-fixed, paraffin embedded (mouse adrenal gland); 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 (CD130) Polyclonal Antibody, Unconjugated (bs-1459R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat adrenal gland); 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 (CD130) Polyclonal Antibody, Unconjugated (bs-1459R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue/cell: Human endometrium 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-IL-6R Beta/CD130/gp130 Polyclonal Antibody, Unconjugated(bs-1459R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: human colon carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-IL-6R Beta/CD130/gp130 Polyclonal Antibody, Unconjugated(bs-1459R) 1:300, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, FITC conjugated (bs-0295G-FITC)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei

Blank control: Raji(blue).
Primary Antibody:Rabbit Anti-CD34 antibody(bs-1459R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min). Antibody (bs-1459R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of bs-1459R at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.