Anti-Pepsin antibody [EPR28182-66] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pepsin antibody [EPR28182-66] - BSA and Azide free (AB312853)

This data was developed using ab312852, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling Pepsin with ab312852 at 1/5000 (0.105 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human colon. The section was incubated with ab312852 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pepsin antibody [EPR28182-66] - BSA and Azide free (AB312853)

This data was developed using ab312852, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded (A) HEK-293T cells tissue labeling Pepsin with ab312852 at 1/1000 (0.528 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T cells transfected with a human PGA3 expression vector containing a his tag, (C) HEK-293T cells transfected with a human PGA4 expression vector containing a his tag, and (D) HEK-293T cells transfected with a human PGA5 expression vector containing a his tag, no staining on (B) HEK-293T cells transfected with a human PGC expression vector containing a his tag and (E) HEK-293T cells transfected with empty vector containing a his tag. The section was incubated with ab312852 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pepsin antibody [EPR28182-66] - BSA and Azide free (AB312853)

This data was developed using ab312852, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Pepsin with ab312852 at 1/5000 (0.105 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human kidney. The section was incubated with ab312852 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pepsin antibody [EPR28182-66] - BSA and Azide free (AB312853)

This data was developed using ab312852, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling Pepsin with ab312852 at 1/5000 (0.105 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human stomach. The section was incubated with ab312852 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-Pepsin antibody [EPR28182-66] - BSA and Azide free (AB312853)

This data was developed using ab312852, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : kidney, skeletal muscle. The molecular weight observed is consistent with what has been described in the reference (PMID : 33620602). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 10 seconds

All lanes:

Western blot - Anti-Pepsin antibody [EPR28182-66] (<a href='/en-us/products/primary-antibodies/pepsin-antibody-epr28182-66-ab312852'>ab312852</a>) at 1/1000 dilution

Lane 1:

Human stomach tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 42 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Pepsin antibody [EPR28182-66] - BSA and Azide free (AB312853)

This data was developed using ab312852, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody demonstrates cross reactivity with human Pepsin A-3, Pepsin A-4 and Pepsin A-5. An additional band with unknown identity at 100 kDa was observed. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. Anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane 1-4 : 180 seconds; Lane 5 : 10 seconds.

All lanes:

Western blot - Anti-Pepsin antibody [EPR28182-66] (<a href='/en-us/products/primary-antibodies/pepsin-antibody-epr28182-66-ab312852'>ab312852</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containi a His-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a Human Pepsin A-3 expression vector containi a His-tag whole cell lysate at 2 µg

Lane 3:

293T cells transfected with a Human PGC expression vector containi a His-tag whole cell lysate at 2 µg

Lane 4:

293T cells transfected with a Human Pepsin A-4 expression vector containi a His-tag whole cell lysate at 10 µg

Lane 5:

Purified Pepsin A-5 recombinant protein (aa63-388) at 20 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 42 kDa

false