Recombinant Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

This data was developed using ab108252, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab181602 was used as a GAPDH loading control.

Two bands observed by ab108252 corresponding to glycosylation and non-glycosylation forms.

Signal in heart tissue is low, we recommend loading more amount of lysate or using lower antibody dilution to improve result.

This data was developed using ab108252, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

ab181602 was used as GAPDH loading control.

Exposure time: Lane 1: 7 seconds; Lane 2-8: 180 seconds.

Two bands observed by ab108252 corresponding to glycosylation and non-glycosylation forms.

Signal in mouse and rat tissues are low, we recommend loading more amount of lysate or using lower antibody dilution to improve result.

This data was developed using ab108252, the same antibody clone in a different buffer formulation.

Lanes 1 - 4: Merged signal (red and green). Green - ab108252 observed at 130 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab108252 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 3% milk in TBS-T (0.1% Tween^®) before incubation with ab108252 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution 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.

This data was developed using ab108252, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labeling ACE2 with ab108252 at 1/6400 dilution. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Staining was visualised using Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

The section was incubated with ab108252 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation (ab108252).

Lanes 1 - 4: Merged signal (red and green). Green - ab108252 observed at 125 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab108252 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 ab108252 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution 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.

This data was developed using ab108252, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labeling ACE2 with ab108252 at 1/6400 dilution. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Staining was visualised using Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

The section was incubated with ab108252 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

This data was developed using ab108252, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labeling ACE2 with ab108252 at 1/6400 dilution. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Staining was visualised using Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

The section was incubated with ab108252 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

ab239924 Immunoprecipitating ACE2 in human testis tissue lysate. 0.35 mg of tissue lysate was incubated with 2 μg primary antibody (1/50). For western blotting a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/1000) was used to confirm successful immunoprecipitation.

Exposure time: 1 second.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.