Anti-CD73 antibody [4G6E3]

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD73 antibody [4G6E3] (AB202122)

Flow cytometric analysis of A431 cells using ab202122 at 1/200 (green) and negative control (red).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (AB202122)

Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling CD73 with ab202122 at 1/200 with DAB staining.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (AB202122)

Immunohistochemical analysis of paraffin-embedded Human esophageal cancer tissue labeling CD73 with ab202122 at 1/200 with DAB staining.

• WB

Lab

Western blot - Anti-CD73 antibody [4G6E3] (AB202122)

False colour image of Western blot : Anti-CD73 antibody [4G6E3] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab202122 was shown to bind specifically to CD73. A band was observed at 65-70 kDa in wild-type A-375 cell lysates with no signal observed at this size in NT5E knockout cell line ab281591 (knockout cell lysate ab282943). A cross-reactive band is observed at 65 kDa in all samples. To generate this image, wild-type and NT5E knockout A-375 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

All lanes:

Western blot - Anti-CD73 antibody [4G6E3] (ab202122) at 1/500 dilution

Lane 1:

Wild-type A-375 cell lysate at 20 µg

Lane 2:

NT5E knockout A-375 cell lysate at 20 µg

Lane 2:

Western blot - Human NT5E knockout A375 cell line (<a href='/en-us/products/cell-lines/human-nt5e-knockout-a375-cell-line-ab281591'>ab281591</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 63 kDa

Observed band size: 65 kDa,65-70 kDa

false

• WB

Supplier Data

Western blot - Anti-CD73 antibody [4G6E3] (AB202122)

All lanes:

Western blot - Anti-CD73 antibody [4G6E3] (ab202122) at 1/500 dilution

All lanes:

A431 cell lysate

Predicted band size: 63 kDa

false

• WB

Lab

Western blot - Anti-CD73 antibody [4G6E3] (AB202122)

Lanes 1 - 3 : Merged signal (red and green). Green - ab202122 observed at 63 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab202122 was shown to recognize NT5E in wild-type A-431 cells as signal was lost at the expected MW in NT5E knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NT5E knockout samples were subjected to SDS-PAGE. ab202122 and ab181602 (Rabbit 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-Mouse IgG H&L (IRDye^® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD73 antibody [4G6E3] (ab202122) at 1/1000 dilution

Lane 1:

Wild-type A-431 whole cell lysate at 20 µg

Lane 2:

NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NT5E knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-nt5e-knockout-a-431-cell-line-ab261895'>ab261895</a>)

Lane 3:

A375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Predicted band size: 63 kDa

false

• WB

Supplier Data

Western blot - Anti-CD73 antibody [4G6E3] (AB202122)

Expected MW is 26.6 kDa

All lanes:

Western blot - Anti-CD73 antibody [4G6E3] (ab202122) at 1/500 dilution

All lanes:

Human CD73 (AA: 30-250) recombinant protein

Predicted band size: 63 kDa

false