Anti-CD147 antibody [10E10] (ab230921)

Formalin-fixed, paraffin-embedded human brain tissue stained for CD147 using ab230921 at 10 μg/ml in immunohistochemical analysis.

Lanes 1 - 4: Merged signal (red and green). Green - ab230921 observed at 42-70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

ab230921 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab230921 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 500 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with ab230921 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab230921) (1x10⁶ in 100μl at 5 μg/ml) for 30 min at 4°C.

The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (ab150117) was used at 1/2000 for 30 min at 4°C.

Isotype control antibody was mouse IgG1κ (ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Formalin-fixed, paraffin-embedded human placenta tissue stained for CD147 using ab230921 at 10 μg/ml in immunohistochemical analysis.

HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for CD147 (green) using ab230921 at 1/100 dilution in ICC/IF.

Flow cytometric analysis of HeLa (human T cell leukemia cell line from peripheral blood) cell line labeling CD147 with ab230921 at 1/100 dilution (red) compared to a nonspecific negative control antibody (blue).

Flow cytometric analysis of Jurkat (human T cell leukemia cell line from peripheral blood) cell line labeling CD147 with ab230921 at 1/100 dilution (red) compared to a nonspecific negative control antibody (blue).