[IF=7.59] Zhen Han. et al. Blockade of the Immune Checkpoint CD47 by TTI-621 Potentiates the Response to Anti-PD-L1 in Cutaneous T Cell Lymphoma. J INVEST DERMATOL. 2023 Mar;:  IF ;  Human.  

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[IF=3.747] Zhao Juan. et al. IRAK4 inhibition: an effective strategy for immunomodulating peri-implant osseointegration via reciprocally-shifted polarization in the monocyte-macrophage lineage cells. BMC ORAL HEALTH. 2023 Dec;23(1):1-17  WB ;  Mouse.  

PubMed:37158847

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

Sample:
Lane 1: Mouse Spleen tissue lysates
Lane 2: Rat Lung tissue lysates
Primary: Anti-CD163 (bsm-54015R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 130 kDa
Observed band size: 170 kDa

Sample:
Lane 1: human thymus tissue lysates
Primary: Anti-CD163 (bsm-54015R) at 1:500 dilution
Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution
Predicted band size: 130 kD
Observed band size: 150 kD

Tissue: Mouse liver
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: 1:500
Primary Ab incubation condition: 1 hour at
room temperature
Secondary Ab: Anti-Rabbit and Mouse
Polymer HRP (Ready to use)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal for
bsm-54015R

Tissue: Human placenta
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: 1:500
Primary Ab incubation condition: 1 hour at
room temperature
Secondary Ab: Anti-Rabbit and Mouse
Polymer HRP (Ready to use)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal
for
bsm-54015R

Tissue: Rat liver
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: 1:500
Primary Ab incubation condition: 1 hour at
room temperature
Secondary Ab: Anti-Rabbit and Mouse
Polymer HRP (Ready to use)
Counter stain: Hematoxylin (Blue)
Comment: Color brown is the positive signal
for
bsm-54015R

Tissue: Human liver
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: 1:500
Primary Ab incubation condition: 1 hour at
room temperature
Secondary Ab: Anti-Rabbit and Mouse
Polymer HRP (Ready to use)
Counter stain:
Hematoxylin (Blue)
Comment: Color brown is the positive signal
for bsm-54015R

Paraformaldehyde-fixed, paraffin embedded (Human placenta); 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 (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) 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 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; Antibody incubation with (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) 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 colon carcinoma); 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 (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) 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 lung carcinoma); 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 (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) 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 spleen tissue); 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 (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human placenta tissue); 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 (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human liver tissue); 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 (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) at 1:50 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human tonsil); 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 (CD163) Monoclonal Antibody, Unconjugated (bsm-54015R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Blank control:THP-1.
Primary Antibody (green line): Rabbit Anti-CD163 antibody (bsm-54015R)
Dilution: 1:50;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1:1000.
Protocol
The cells were 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.