Mannose Receptor(CD206) Recombinant Rabbit Monoclonal Antibody [JF0953] - BSA and Azide free

Western blot analysis of Mannose Receptor(CD206) on different lysates with Rabbit anti-Mannose Receptor(CD206) antibody (HA750332) at 1/5,000 dilution.

Lane 1: MOLT-4 cell lysate (20 µg/Lane)
Lane 2: Mouse lung tissue lysate (40 µg/Lane)

Predicted band size: 166 kDa
Observed band size: 166 kDa

Exposure time: 18 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750332) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

All lanes: Western blot analysis of CD206 with anti-CD206 antibody[JF0953] (HA750332) at 1:1,000 dilution.
Lane 1: Wild-type HepG2 whole cell lysate (10 µg).
Lane 2: CD206 knockdown HepG2 whole cell lysate (10 µg).
Lane 3: CD206 knockdown HepG2 whole cell lysate (10 µg).

ET1702-04 was shown to specifically react with CD206 in wild-type Hela cells. Weakened bands were observed when CD206 knockdown samples were tested. Wild-type and CD206 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1702-04, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Mannose Receptor(CD206) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA750332, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Human lung tissue lysate
Lane 2: HepG2 cell lysate

Western blot analysis of Mannose Receptor(CD206) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA750332, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Mouse lung tissue lysate
Lane 2: D3 cell lysate

Immunocytochemistry analysis of HepG2 cells labeling Mannose Receptor(CD206) with Rabbit anti-Mannose Receptor(CD206) antibody (HA750332) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Mannose Receptor(CD206) antibody (HA750332) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of HepG2 cells labeling Mannose Receptor(CD206).

Cells were fixed and permeabilized. Then stained with the primary antibody (HA750332, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).