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☑ Cell treatment (CT)
Western blot analysis of Mannose Receptor(CD206) on different lysates with Rabbit anti-Mannose Receptor(CD206) antibody (HA751919) 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: HA751919, 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
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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: HA751919, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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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: HA751919, 1/3,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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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: HA751919, 1/3,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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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: HA751919, 1/1,000, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
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☑ Cell treatment (CT)
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 HA751919 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751919 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) 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: HA751919 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 HA751919 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: HA751919, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 166 kDa
Observed band size: 250 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"