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

4 product images

• 

  Flow Cytometry (Intracellular) - Anti-M6PR (cation independent) antibody [MEM-238] - Lysosome Membrane Marker (ab8093)

  M6PR (cation independent) Flow Cytometry (Intracellular) staining using mouse Anti-M6PR (cation independent) antibody

  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 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  

• 

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

  M6PR (cation independent) Western blot staining using mouse Anti-M6PR (cation independent) antibody

  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

• 

  Flow Cytometry - Anti-M6PR (cation independent) antibody [MEM-238] (ab8093)

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

• 

  Flow Cytometry - Anti-M6PR (cation independent) antibody [MEM-238] (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.