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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling ACE2 with ab108252 at a concentration of 0.01µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32 mins. ab108252 Anti-ACE2 antibody [EPR4435(2)] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling ACE2 with ab108252 at a concentration of 0.01µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32 mins. ab108252 Anti-ACE2 antibody [EPR4435(2)] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

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.

• IP

Lab

Immunoprecipitation - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (AB239924)

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.

All lanes:

Immunoprecipitation - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924) at 1/500 dilution

Lane 1:

Human testis tissue lysate at 10 µg

Lane 2:

ab239924 + Human testis tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab239924 in Human testis tissue lysate

Predicted band size: 92 kDa

Observed band size: 110 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

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.

• WB

Lab

Western blot - 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.

All lanes:

Western blot - Anti-ACE2 antibody [EPR4435(2)] (<a href='/en-us/products/primary-antibodies/ace2-antibody-epr44352-ab108252'>ab108252</a>) at 1/1000 dilution

Lane 1:

Human heart tissue lysate at 20 µg

Lane 2:

Rat heart tissue lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 92 kDa

Observed band size: 110 kDa,120 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (AB239924)

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.

All lanes:

Western blot - Anti-ACE2 antibody [EPR4435(2)] (<a href='/en-us/products/primary-antibodies/ace2-antibody-epr44352-ab108252'>ab108252</a>) at 1/1000 dilution

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: 104 kDa,60 kDa,76 kDa,92 kDa

Observed band size: 125 kDa,75 kDa

false

• WB

Lab

Western blot - 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.

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.

All lanes:

Western blot - Anti-ACE2 antibody [EPR4435(2)] (<a href='/en-us/products/primary-antibodies/ace2-antibody-epr44352-ab108252'>ab108252</a>) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 30 µg

Lane 2:

ACE2 knockout HepG2 cell lysate at 30 µg

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

Lab

Western blot - 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. Different batches of ab108252 were tested on Human kidney lysate at 0.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 120 kDa.

All lanes:

Western blot - Anti-ACE2 antibody [EPR4435(2)] (<a href='/en-us/products/primary-antibodies/ace2-antibody-epr44352-ab108252'>ab108252</a>)

Predicted band size: 92 kDa

false

• WB

Lab

Western blot - 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 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.

All lanes:

Western blot - Anti-ACE2 antibody [EPR4435(2)] (<a href='/en-us/products/primary-antibodies/ace2-antibody-epr44352-ab108252'>ab108252</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate

Lane 4:

Mouse spleen tissue lysate

Lane 5:

Mouse lung tissue lysate

Lane 6:

Rat testis tissue lysate

Lane 7:

Rat spleen tissue lysate

Lane 8:

Rat lung tissue lysate

Secondary

All lanes:

Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 92 kDa

Observed band size: 110 kDa,120 kDa

false

• WB

Unknown

Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (AB239924)

All lanes:

Western blot - Anti-ACE2 antibody [EPR4435(2)] (<a href='/en-us/products/primary-antibodies/ace2-antibody-epr44352-ab108252'>ab108252</a>) at 1/1000 dilution

Lane 1:

Human fetal kidney lysate at 10 µg

Lane 2:

Human testis lysate at 10 µg

Predicted band size: 92 kDa

false