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Immunohistochemical analysis of paraffin-embedded human B lymphocyte tumor tissue with Mouse anti-CD43 antibody (EM1901-70) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-70) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-CD43 antibody (EM1901-70) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-70) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of CD43 was done on HL-60 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-70, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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Western blot analysis of CD43 on different lysates with Rabbit anti-CD43 antibody (EM1901-70) at 1/1,000 dilution.
Lane 1: HL-60 (Human acute promyelocytic leukemia cells) cell lysat
Lane 2: THP-1 (Human acute monoblastic leukemia cell) cell lysate
Lane 3: U-937 (Human acute monocytic leukemia cells) cell lysate
Lane 4: HT-29 (Human colorectal adenocarcinoma cell) cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 25 seconds; ECL: K1802
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: EM1901-70, 1/1,000 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: 40 kDa
Observed band size: 100-130 kDa
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