Anti-CLPTM1 antibody [EPR8800] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CLPTM1 antibody [EPR8800] - BSA and Azide free (AB248641)

This data was developed using ab133756, the same antibody clone in a different buffer formulation.

Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling CLPTM1 with purified ab133756 at 1/200 dilution (1 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

Lab

Western blot - Anti-CLPTM1 antibody [EPR8800] - BSA and Azide free (AB248641)

Lanes 1 - 4 : Merged signal (red and green). Green - ab133756 observed at 76 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab133756 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in CLPTM1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and CLPTM1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3pc Milk. ab133756 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 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 (ab133756).

All lanes:

Western blot - Anti-CLPTM1 antibody [EPR8800] (<a href='/en-us/products/primary-antibodies/clptm1-antibody-epr8800-ab133756'>ab133756</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

CLPTM1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

A549 whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Predicted band size: 76 kDa

Observed band size: 76 kDa

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