TLR6 Rabbit pAb(bs-2716R)-50ul

Rat splenocytes stained with Anti- TLR6/CD286 Polyclonal Antibody, PE-CY5.5 Conjugated (bs-2716R-PE-Cy5.5) at 1:25.

P‌‌araformaldehyde-fixed, paraffin embedded (rat spleen tissue); 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 (TLR6) Polyclonal Antibody, Unconjugated (bs-2716R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.

Blank control: Raji (blue).
Primary Antibody:Rabbit Anti-TLR6 antibody(bs-2716R), Dilution: 0.2μg 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) and then permeabilized with ice-cold 90% methanol for 30 min on ice.The cells were washed twice with phosphate-buffered saline (PBS).The cells were then 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 antibody (bs-2716R, 0.2μg /1x10^6 cells) for 30 min on ice. The secondary antibody used was Goat Anti-rabbit IgG/PE antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.

Tissue/cell: human lung 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-TLR6 Polyclonal Antibody, Unconjugated(bs-2716R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control:HUVEC.
Primary Antibody (green line): Rabbit Anti-TLR6 antibody (bs-2716R)
Dilution: 1ug/Test;
Secondary Antibody (white blue line) : Goat anti-rabbit IgG-AF488
Dilution: 0.5ug/Test.
Isotype control（orange line）：Normal Rabbit IgG
Protocol
The cells were 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.