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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; Incubation: Anti-ID1 Antibody, conjugated (SL3529R-FITC) 1:400, 1.5 hours at 37°C; DAPI (5ug/ml, blue, SLC0033) was used to stain the cell nuclei.
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; Incubation: Anti-ID1 Antibody, conjugated (SL3529R-FITC) 1:400, 1.5 hours at 37°C; DAPI (5ug/ml, blue, SLC0033) was used to stain the cell nuclei.
Blank control: mouse spleen cells(blue). Primary Antibody: Rabbit Anti- ID1/FITC Conjugated antibody (SL3529R /FITC, green line), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG/FITC(orange) ,used under the same conditions.
Protocol
The cells were fixed with 4% 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 (SL3529R/FITC, 1μg /1x10^6 cells) for 30 min on ice. Acquisition of 20,000 events was performed.
Blank control: mouse kidney cells(blue).
Primary Antibody: Rabbit Anti- ID1/FITC Conjugated antibody (SL3529R /FITC, green line), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG/FITC(orange) ,used under the same conditions.
Protocol
The cells were fixed with 4% 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 (SL3529R/FITC, 1μg /1x10^6 cells) for 30 min on ice. Acquisition of 20,000 events was performed.
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