|
|
WB on human PMN (peripheral blood mononuclear cells isolated from buffycoat; denatured, reduced) using Rabbit antibody to Pr3 (OSP00050W) at 1: 500 dilution; blocked with 1% LFDM for 15 minutes at room temperature with shake, primary antibody incubated for 15 minutes at room temperature, washed 3 times with PBST, 5 minutes each. Secondary antibody was also incubated for 15 minutes at room temperature. |
|
Human PBMC were isolated and adjusted to 106 cells mL-1. Cells were fixed with 2% formaldehyde for 10 minutes at 37°C. Washed twice with PBS before cytospin the cells onto microscope slides. Cells were blocked with PBS containing 1%BSA for 20 minutes at room temperature (RT). Excess of blocking solution was removed and cells were then incubated with Rabbit antibody to Pr3 (OSP00050W) for 30 min at RT (diluted 1:100 in the blocking buffer). Washed 3X with PBS and incubated with anti-Rabbit Alexa 586 for further 30 minutes. Washed as before and nuclear counterstained with DAPI. Neutrophils and Monocytes, known to have PR3 are intensely stained by the Rabbit antibody to c-terminal region of Pr3 (Wegener autoantigen): whole serum (OSP00050W). |
|
Human PBMC were isolated and adjusted to 106 cells mL-1. Cells were fixed with 2% formaldehyde for 10 minutes at 37°C. Washed twice with PBS before cytospin the cells onto microscope slides. Cells were blocked with PBS containing 1%BSA for 20 minutes at room temperature (RT). Excess of blocking solution was removed and cells were then incubated with Rabbit antibody to Pr3 (OSP00050W) for 30 min at RT (diluted 1:100 in the blocking buffer). Washed 3X with PBS and incubated with anti-Rabbit Alexa 586 for further 30 minutes. Washed as before and nuclear counterstained with DAPI. Neutrophils and Monocytes, known to have PR3 are intensely stained by the Rabbit antibody to c-terminal region of Pr3 (Wegener autoantigen): whole serum (OSP00050W). |
|
Human PBMC were isolated and adjusted to 106 cells mL-1. Cells were fixed with 2% formaldehyde for 10 minutes at 37°C. Washed twice with PBS before cytospin the cells onto microscope slides. Cells were blocked with PBS containing 1%BSA for 20 minutes at room temperature (RT). Excess of blocking solution was removed and cells were then incubated with Rabbit antibody to Pr3 (OSP00050W) for 30 min at RT (diluted 1:100 in the blocking buffer). Washed 3X with PBS and incubated with anti-Rabbit Alexa 586 for further 30 minutes. Washed as before and nuclear counterstained with DAPI. Neutrophils and Monocytes, known to have PR3 are intensely stained by the Rabbit antibody to c-terminal region of Pr3 (Wegener autoantigen): whole serum (OSP00050W). |
|
Human PBMC were isolated and adjusted to 106 cells mL-1. Cells were fixed with 2% formaldehyde for 10 minutes at 37°C. Washed twice with PBS before cytospin the cells onto microscope slides. Cells were blocked with PBS containing 1%BSA for 20 minutes at room temperature (RT). Excess of blocking solution was removed and cells were then incubated with Rabbit antibody to Pr3 (OSP00050W) for 30 min at RT (diluted 1:100 in the blocking buffer). Washed 3X with PBS and incubated with anti-Rabbit Alexa 586 for further 30 minutes. Washed as before and nuclear counterstained with DAPI. Neutrophils and Monocytes, known to have PR3 are intensely stained by the Rabbit antibody to c-terminal region of Pr3 (Wegener autoantigen): whole serum (OSP00050W). |