Mannose Receptor(CD206) Recombinant Rabbit Monoclonal Antibody [PSH23-93] - BSA and Azide free

☑ Cell treatment (CT)

This data was developed using HA724408, the same antibody clone in a different buffer formulation.
Western blot analysis of Mannose Receptor(CD206) on different lysates with Rabbit anti-Mannose Receptor(CD206) antibody (HA724408) at 1/5,000 dilution.

Lane 1: THP-1 (Human acute monoblastic leukemia cells) cell lysate
Lane 2: THP-1 treated with 20ng/ml PMA for 24 hours then treated with 20ng/ml IL-4 for 48 hours cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 20 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724408, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 166 kDa
Observed band size: 180 kDa

This data was developed using HA724408, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Colon
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724408, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724408, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Liver
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724408, 1/3,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724408, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Lung
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724408, 1/3,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.

This data was developed using HA724408, the same antibody clone in a different buffer formulation.
Application: Immunofluorescence (IF-tissue)

Species: Human
Tissue: Liver
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724408, 1/1,000, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.

☑ Cell treatment (CT)

This data was developed using HA724408, the same antibody clone in a different buffer formulation.
Immunoprecipitation (IP)

Mannose Receptor(CD206) was immunoprecipitated in 0.2 mg THP-1 (Human acute monoblastic leukemia cell) treated with 20ng/mL TPA for 24 hours then treated with 20ng/mL IL-4 for 48 hours cell lysate with HA724408 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724408 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: THP-1 treated with 20ng/mL TPA for 24 hours then treated with 20ng/mL IL-4 for 48 hours cell lysate
Lane 2: HA724408 IP in THP-1 treated with 20ng/mL TPA for 24 hours then treated with 20ng/mL IL-4 for 48 hours cell lysate
Lane 3: Rabbit IgG instead of HA724408 in THP-1 treated with 20ng/mL TPA for 24 hours then treated with 20ng/mL IL-4 for 48 hours cell lysate

Exposure time: 3 minutes
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724408, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 166 kDa
Observed band size: 250 kDa