| 亚型: IgG | 验证方法: 1. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
2. Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
3. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
4. Paraformaldehyde-fixed, paraffin embedded (human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
5. Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (GFAP) Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
6. Blank control: U87MG.
Primary Antibody (green line): Rabbit Anti-GFAP antibody (TMAB-00762)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. 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.
7. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with GFAP Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with GFAP Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with GFAP Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Rat Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with GFAP Polyclonal Antibody, Unconjugated (TMAB-00762) at 1:200 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining. |