Recombinant Anti-M6PR (cation dependent) antibody [EPR7691] - BSA and Azide free (ab236067)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Empty
Lane 3: M6PR knockout HAP1 whole cell lysate (20 µg)
Lane 4: A549 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab134153 observed at 46 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab134153 was shown to specifically react with M6PR when M6PR knockout samples were used. Wild-type and M6PR knockout samples were subjected to SDS-PAGE. ab134153 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134153).

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling M6PR (cation dependent) with Purified ab134153 at 1:100 dilution (8.6 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134153).

Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling M6PR (cation dependent) with purified ab134153 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134153).

Equilibrium disassociation constant (K_D)
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134153).