Anti-ACE2 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody (AB65863)

IHC image of ACE2 staining in human kidney carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab65683, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• WB

Lab

Western blot - Anti-ACE2 antibody (AB65863)

Lanes 1 - 4 : Merged signal (red and green). Green - ab65863 observed at 125 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab65863 was shown to react with ACE2 in Caco-2 wild-type cells in western blot with loss of signal observed in ACE2 knockout cell line ab273731 (knockout cell lysate ab275516). Wild-type and ACE2 knockout Caco-2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab65863 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ACE2 antibody (ab65863) at 1 µg/mL

Lane 1:

Wild-type Caco-2 cell lysate at 30 µg

Lane 2:

ACE2 knockout Caco-2 cell lysate at 30 µg

Lane 2:

Western blot - Human ACE2 knockout Caco-2 cell line (<a href='/en-us/products/cell-lines/human-ace2-knockout-caco-2-cell-line-ab273731'>ab273731</a>)

Lane 3:

Calu-3 cell lysate at 30 µg

Lane 4:

A549 cell lysate at 30 µg

Predicted band size: 92 kDa

Observed band size: 125 kDa

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• WB

Lab

Western blot - Anti-ACE2 antibody (AB65863)

Lanes 1 - 4 : Merged signal (red and green). Green - ab65863 observed at 130 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab65863 was shown to react with ACE2 in wild-type HepG2 cells in western blot with loss of signal observed in ACE2 knockout cell line ab273733 (knockout cell lysate ab275495). Wild-type and ACE2 knockout HepG2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab65863 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ACE2 antibody (ab65863) at 1 µg/mL

Lane 1:

Wild-type HepG2 cell lysate at 30 µg

Lane 2:

ACE2 knockout HepG2 cell lysate at 30 µg

Lane 2:

Western blot - Human ACE2 knockout Hep G2 cell line (<a href='/en-us/products/cell-lines/human-ace2-knockout-hep-g2-cell-line-ab273733'>ab273733</a>)

Lane 3:

Calu-3 cell lysate at 30 µg

Lane 4:

A549 cell lysate at 30 µg

Predicted band size: 92 kDa

Observed band size: 130 kDa

false

• WB

Unknown

Western blot - Anti-ACE2 antibody (AB65863)

ACE2 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-ACE2 antibody (ab65863) at 1 µg/mL

All lanes:

Human kidney tissue lysate - total protein (<a href='/en-us/products/unavailable/human-kidney-tissue-lysate-total-protein-ab30203'>ab30203</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 92 kDa

Observed band size: 110 kDa

true