VALIDATION IMAGES
Blocking buffer: 5% NFDM/TBST
Primary ab dilution: 1:1000
Primary ab incubation condition: 2 hours at room temperature
Secondary ab: Goat Anti-Rabbit IgG H&L (HRP)
Lysate: HepG2, 4T1, SH-SY5Y, BRL
Protein loading quantity: 20 μg
Exposure time: 3 s
Predicted MW: 50 kDa
Observed MW: 50 kDa
Tissue: Rat kidney
Section type: Formalin fixed & Paraffin -embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary ab dilution: 1:100
Primary ab incubation condition: 1 hour at room temperature
Secondary ab: SP Kit(Mouse)(sp-0024)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal for bsm-60482M
Tissue: Mouse heart
Section type: Formalin fixed & Paraffin -embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary ab dilution: 1:100
Primary ab incubation condition: 1 hour at room temperature
Secondary ab: SP Kit(Mouse)(sp-0024)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal for bsm-60482M
Tissue: Human skeletal muscle
Section type: Formalin fixed & Paraffin -embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0 Primary ab dilution: 1:100
Primary ab incubation condition: 1 hour at room temperature
Secondary ab: SP Kit(Mouse)(sp-0024)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal for bsm-60482M
25 ug total protein per lane of various lysates (see on figure) probed with ATP5A1 monoclonal antibody, unconjugated (bsm-60482M) at 1:500 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Skeletal muscle; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with ATP5A1 Monoclonal Antibody, Unconjugated (bsm-60482M) at 1:100 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.