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Sample: Lane 1: Human Raji cell Lysates Primary: Anti-CD79b (bsm-60240R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 23kDa Observed band size: 35kDa

25 ug total protein per lane of various lysates (see on figure) probed with CD79b monoclonal antibody, unconjugated (bsm-60240R) at 1:2000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Paraformaldehyde-fixed, paraffin embedded (rat spleen); 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 (CD79b) Monoclonal Antibody, Unconjugated (bsm-60240R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Tissue: Human cervical carcinoma 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: 1:1000 Primary ab incubation condition: 1 hour at room temperature Counter stain: Hematoxylin Comment: Color brown is the positive signal for bsm-60240R

Tissue: Human spleen 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: 1:1000 Primary ab incubation condition: 1 hour at room temperature Counter stain: Hematoxylin Comment: Color brown is the positive signal for bsm-60240R

Tissue: Rat spleen 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: 1:1000 Primary ab incubation condition: 1 hour at room temperature Counter stain: Hematoxylin Comment: Color brown is the positive signal for bsm-60240R

Tissue: Mouse spleen 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: 1:1000 Primary ab incubation condition: 1 hour at room temperature Counter stain: Hematoxylin Comment: Color brown is the positive signal for bsm-60240R

4% Paraformaldehyde-fixed Ramos (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (CD79b) monoclonal Antibody, unconjugated (bsm-60240R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

The Ramos(H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Primary Antibody (green): Rabbit Anti-CD79b antibody (bsm-60240R): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-60295G-FITC): 1 μg/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.