Anti-M6PR (cation independent) antibody [MEM-238] - Lysosome Membrane Marker

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-M6PR (cation independent) antibody [MEM-238] - Lysosome Membrane Marker (AB8093)

Flow cytometry surface staining analysis of human peripheral whole blood showing separation of human neutrophil granulocytes (red) from lymphocytes black using ab8093 at 2 μg/ml GAM APC.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-M6PR (cation independent) antibody [MEM-238] - Lysosome Membrane Marker (AB8093)

Flow cytometry analysis of human peripheral whole blood stained using ab8093 in 2 μg/ml and GAM APC.

• WB

Supplier Data

Western blot - Anti-M6PR (cation independent) antibody [MEM-238] - Lysosome Membrane Marker (AB8093)

Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of Jurkat, K562, Raji, and HeLa cell lines, mixed and heated (100°C, 5 min) with reducing and non-reducing SDS-loading buffer. Samples were resolved using 7% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed with ab8093 followed by IRDye 800CW Goat-anti-Mouse IgG (green). Mouse anti-GAPDH monoclonal antibody FF26A conjugated with DyLight 680 (0.1 µg/ml), was used as the loading control (red). Multiplex fluorescent Western blot detection was performed. CD222 molecules were detected at ~250 kDa in all analysed cell lines.

All lanes:

Western blot - Anti-M6PR (cation independent) antibody [MEM-238] - Lysosome Membrane Marker (ab8093)

Lane 1:

Jurkat whole cell extract, with reducing SDS loading buffer

Lane 2:

K562 whole cell extract, with reducing SDS loading buffer

Lane 3:

Raji whole cell extract, with reducing SDS loading buffer

Lane 4:

HeLa whole cell extract, with reducing SDS loading buffer

Lane 5:

Jurkat whole cell extract, with non-reducing SDS loading buffer

Lane 6:

K562 whole cell extract, with non-reducing SDS loading buffer

Lane 7:

Raji whole cell extract, with non-reducing SDS loading buffer

Lane 8:

HeLa whole cell extract, with non-reducing SDS loading buffer

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• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-M6PR (cation independent) antibody [MEM-238] - Lysosome Membrane Marker (AB8093)

Overlay histogram showing HeLa cells stained with ab8093 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8093, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.