VALIDATION IMAGES
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:5000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: 1: Mouse liver, 2: Rat spleen
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 121 kDa
Observed MW: 150 kDa
Human thymus tissue lysate probed with CD163 (8G1) Monoclonal Antibody (bsm-54015R)at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation for 60 min at 37˚C.
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:5000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: 1: THP-1 (Negative control), 2: Human
liver
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 121 kDa
Observed MW: 150 kDa
Flow cytometric analysis of thp-1 cells with CD163 (8G1) Monoclonal Antibody (bsm-54015R) at a 1:50 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:5000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: 1: THP-1 (Negative control), 2: Human
liver
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 121 kDa
Observed MW: 150 kDa
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:5000
Primary Ab incubation condition: 2 hours at
room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: 1: Mouse liver, 2: Rat spleen
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 121 kDa
Observed MW: 150 kDa
Paraformaldehyde-fixed, paraffin embedded Mouse Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-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; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-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; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with CD163 Monoclonal Antibody, Unconjugated(bsm-54015R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Lane 1: Mouse Spleen tissue lysates; Lane 2: Rat Lung tissue lysates probed with CD163 Monoclonal Antibody, Unconjugated (bsm-54015R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
25 ug total protein per lane of various lysates (see on figure) probed with CD163 monoclonal antibody, unconjugated (bsm-54015R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.