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☑ Relative expression (RE)
Western blot analysis of CD27 on different lysates with Rabbit anti-CD27 antibody (HA751926) at 1/10,000 dilution.
Lane 1: Raji (Human Burkitt's lymphoma cells) cell lysate
Lane 2: MCF7 (Human breast cancer cells) cell lysate
Lane 3: Ramos (Human Burkitt's lymphoma cells) cell lysate
Lane 4: Daudi (Human Burkitt's lymphoma cells) cell lysate
Lane 5: THP-1 (Human acute monoblastic leukemia cells) cell lysate
Lysates/proteins at 10 µg/Lane.
Exposure time: 12 seconds; ECL: K1801
MCF7 and THP-1 are negative controls (PMID: 11062504).
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751926, 1/10,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 29 kDa
Observed band size: 50 kDa
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☑ Relative expression (RE)
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: Raji (Human Burkitt's lymphoma cells) / MCF7 (Human breast cancer cells)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA751926, 1/2,000, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
Negative control: MCF7 (PMID: 11062504).
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Tonsil
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: HA751926, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Lymph nodes
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: HA751926, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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☑ Relative expression (RE)
Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Skeletal muscle
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: HA751926, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
Negative expression of CD27 in human skeletal muscle is consistent with the predicted expression pattern.
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☑ Relative expression (RE)
Application: Flow Cytometry (Intra)
Species: Human
Sample: Raji (Human Burkitt's lymphoma cells) / MCF7 (Human breast cancer cells)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA751926 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Negative control: MCF7 (PMID: 11062504).
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