Anti-Podoplanin (MAP tag) [PMab-1]

ELISA using PMab-1 and Multi Tag protein 2. Binding curves of the mouse IgG1 chimeric version of the anti-MAP antibody PMab-1 (red line) and isotype control (orb256381, anti-fluorescein; orange line) to an ELISA plate coated with Multi Tag protein 2 at a concentration of 5 ug/ml.

Flow-cytometry using the anti-Podoplanin (MAP tag) antibody PMab-1 NIH3T3 cells were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the rabbit IgG-chimeric version of PMab-1 (orb758777, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Immunofluorescence staining of HEK293 cells with Anti-Podoplanin (MAP tag) PMab-1. Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells transfected with Podoplanin (MAP tag) expressing plasmid (A) and non-transfected HEK293 cells (B), permeabilized with 0.15% Triton stained with the chimeric rabbit IgG version of PMab-1 (orb758777) (1:200 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane and cytoplasmic staining. The nuclear stain is DAPI (blue).

Western Blot using anti-Podoplanin (MAP tag) antibody PMab-1 (orb758777) Mouse thymus (A), mouse brain (B), NIH3T3 cells (C) and mouse kidney (D) lysates (35 µg protein in RIPA buffer) were resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of 1B4 (orb613976) at 0.01 µg/ml, 0.1 µg/ml, 0.001 µg/ml and 0.003 µg/ml, respectively, before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

Western Blot using Anti-Podoplanin (MAP tag) antibody PMab-1. Podoplanin (MAP tag) expressing plasmid transfected (A) and non-transfected (B) HEK293 cells lysate (3 µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of PMab-1 (orb758777) at 0.00001 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.