RGN | Regucalcin Rabbit pAb(bs

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 (Regucalcin) Polyclonal Antibody, Unconjugated (bs-2665R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Sample: Liver (Mouse) Lysate at 40 ug
_x000D_ Primary: Anti-Regucalcin (bs-2665R) at 1/300 dilution
_x000D_ Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
_x000D_ Predicted band size: 33 kD
_x000D_ Observed band size: 33 kD
_x000D_

Blank control（black line）:HepG2.
_x000D_ Primary Antibody (green line): Rabbit Anti-Regucalcin antibody (bs-2665R)
_x000D_ Dilution:2ug/Test;
_x000D_ Secondary Antibody（white blue line）: Goat anti-rabbit IgG-AF488
_x000D_ Dilution: 0.5ug/Test.
_x000D_ Isotype control（orange line）: Normal Rabbit IgG
_x000D_ Protocol
_x000D_ The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, 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.

P‌‌araformaldehyde-fixed, paraffin embedded (human liver); 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; Incubation with (Regucalcin) Polyclonal Antibody, Unconjugated (bs-2665R) 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 kidney); 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; Incubation with (Regucalcin) Polyclonal Antibody, Unconjugated (bs-2665R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

P‌‌araformaldehyde-fixed, paraffin embedded (human kidney); 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; Incubation with (Regucalcin) Polyclonal Antibody, Unconjugated (bs-2665R) 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; Incubation with (Regucalcin) Polyclonal Antibody, Unconjugated (bs-2665R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

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; Incubation with (Regucalcin) Polyclonal Antibody, Unconjugated (bs-2665R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.