Anti-STEAP1 antibody [EPR29701-75]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STEAP1 antibody [EPR29701-75] (AB324263)

Immunohistochemical analysis of paraffin-embedded Human prostate carcinoma tissue labeling STEAP1 with ab324263 at 1/100 (5.27 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human prostate carcinoma (PMID : 10588738).
The primary antibody was incubated for 30 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-STEAP1 antibody [EPR29701-75] (AB324263)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling STEAP1 with ab324263 at 1/100 (5.27 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human colon (PMID : 10588738).
The primary antibody was incubated for 30 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-STEAP1 antibody [EPR29701-75] (AB324263)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling STEAP1 with ab324263 at 1/100 (5.27 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human cerebrum (PMID : 10588738).
The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica BiosystemsBOND® 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STEAP1 antibody [EPR29701-75] (AB324263)

Immunohistochemical analysis of paraffin-embedded HEK-293T (human embryonic kidney epithelial cell) cells labelling STEAP1 with ab324263 at 1 : 8000 (0.066 μg/ml). Positive staining on (A) HEK-293T transfected with a human STEAP1 expression vector containing a His tag, no staining on (B) HEK-293T transfected with a human STEAP1B1 expression vector containing a His tag, (C) HEK-293T transfected with a human STEAP1B2 expression vector containing a His tag, (D) HEK-293T transfected with a human STEAP2 expression vector containing a His tag, (E) HEK-293T transfected with a human STEAP3 expression vector containing a His tag, (F) HEK-293T transfected with a human STEAP4 expression vector containing a His tag, and (G) HEK-293T transfected with empty vector containing a His tag. The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody was a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) kit.

Counterstained with Hematoxylin.

Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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-STEAP1 antibody [EPR29701-75] (AB324263)

Immunohistochemical analysis of paraffin-embedded (A) LNCaP (human prostate carcinoma epithelial cell) cell pellet. (B) PC-3 (human prostate adenocarcinoma epithelial cell) cell pellet. tissue labeling STEAP1 with ab324263 at 1/100 (5.27 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) LNCaP cell pellet, no staining on (B) PC-3 cell pellet.
The primary antibody was incubated for 30 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-STEAP1 antibody [EPR29701-75] (AB324263)

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

Low expression : PC-3, DU 145 (PMID : 37041154).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 37041154).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-STEAP1 antibody [EPR29701-75] (ab324263) at 1/1000 dilution

Lane 1:

22Rv1 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 30 kDa,32 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-STEAP1 antibody [EPR29701-75] (AB324263)

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

This antibody does not cross-react with human STEAP1B1, STEAP1B2, STEAP2, STEAP3, STEAP4.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-STEAP1 antibody [EPR29701-75] (ab324263) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T cells transfected with a human STEAP1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human STEAP1B1 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human STEAP1B2 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 5:

293T cells transfected with a human STEAP2 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 6:

293T cells transfected with a human STEAP3 expression vector containing a His-tag, whole cell lysate at 20 µg

Lane 7:

293T cells transfected with a human STEAP4 expression vector containing a His-tag, whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 30 kDa,32 kDa,15-50 kDa,15 kDa

false

Exposure time: 6s