Anti-Uridine Phosphorylase 1 antibody [EPR29005-505]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse small intestine tissue staining Uridine Phosphorylase 1 with ab325391 at a 1/2000 (0.258 μg/ml) dilution, ab307365 anti-Sodium/Hydrogen Exchanger 3/NHE-3 used at a 1/5000 (0.126 μg/ml) dilution and ab272692 anti-MUC2 used at a 1/2000 (0.26 μg/ml) dilution.

Panel A : anti-Uridine Phosphorylase 1 (green; Opal™690), anti-Sodium/Hydrogen Exchanger 3/NHE-3 (magenta; Opal™520), anti-MUC2 (gray; Opal™570) on mouse small intestine.

Panel B : anti-Uridine Phosphorylase 1 staining cytoplasm and nucleus of epithelium in mouse small intestine.

Panel C : anti-Sodium/Hydrogen Exchanger 3/NHE-3 staining apical and luminal facing edges of surface cells in mouse small intestine.

Panel D : anti-MUC2 staining on goblet cells in mouse small intestine.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325391, ab307365 and ab272692 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

Immunohistochemical analysis of paraffin-embedded (A) ES-D3 (mouse blastocyst-derived embryonic stem cell) cell pellet (B) F9 (mouse embryonal carcinoma epithelial cell) cell pellet (C) EL4 (mouse lymphoma T lymphocyte) cell pellet (D) A20 (mouse reticulum sarcoma B lymphocyte) cell pellet tissue labeling Uridine Phosphorylase 1 with ab325391 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) ES-D3 and (B) F9 cell pellets, no staining on (C) EL4 and (D) A20 cell pellets.
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-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

Immunohistochemical analysis of paraffin-embedded Mouse glioblastoma tissue labeling Uridine Phosphorylase 1 with ab325391 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse glioblastoma.
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-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

Immunohistochemical analysis of paraffin-embedded Mouse small intestine tissue labeling Uridine Phosphorylase 1 with ab325391 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse small intestine (PMID : 14715930).
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-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

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

Negative control : mouse liver, mouse testis, mouse brain (PMID : 14715930), EL4, A20.

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

All lanes:

Western blot - Anti-Uridine Phosphorylase 1 antibody [EPR29005-505] (ab325391) at 1/1000 dilution

Lane 1:

ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate at 20 µg

Lane 2:

EL4 (mouse lymphoma t lymphocyte) whole cell lysate at 20 µg

Lane 3:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate at 20 µg

Lane 5:

Mouse small intestine lysate at 20 µg

Lane 6:

Mouse liver lysate at 20 µg

Lane 7:

Mouse testis lysate at 20 µg

Lane 8:

Mouse brain 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 1/20000 dilution

Observed band size: 34 kDa,36 kDa

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Uridine Phosphorylase 1 with ab325391 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Weak nuclear staining on sporadic cells in mouse testis.
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-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Uridine Phosphorylase 1 with ab325391 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Nearly no staining on mouse liver.
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-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Uridine Phosphorylase 1 with ab325391 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse cerebrum (PMID : 14715930).
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-Uridine Phosphorylase 1 antibody [EPR29005-505] (AB325391)

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

This antibody does not cross-react with overexpressed mouse UPP2 by western blot.

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

All lanes:

Western blot - Anti-Uridine Phosphorylase 1 antibody [EPR29005-505] (ab325391) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 2:

293T cells transfected with mouse Uridine Phosphorylase 1 (UPP1) expression vector containing a myc-His-tag® whole cell lysate at 5 µg

Lane 3:

293T cells transfected with mouse Uridine Phosphorylase 2 (UPP2) expression vector containing a myc-His-tag® whole cell lysate at 5 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: 34 kDa,36 kDa

false

Exposure time: 15s