Anti-PYGO2 antibody [EPR27168-186]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PYGO2 antibody [EPR27168-186] (AB316318)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling PYGO2 with ab316318 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human placenta.
The section was incubated with ab316318 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-PYGO2 antibody [EPR27168-186] (AB316318)

Immunohistochemical analysis of paraffin-embedded Panel A. Human normal stomach Panel B. Human gastric cancer tissue labeling PYGO2 with ab316318 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nearly no staining on human normal stomach (Panel. A), nuclear staining on human gastric cancer (Panel. B) (PMID : 23865714).
The two images come from same donor's tumour and adjacent normal.
The section was incubated with ab316318 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-PYGO2 antibody [EPR27168-186] (AB316318)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling PYGO2 with ab316318 at 1/5000 (0.103 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung cancer (PMID : 23865714)
The section was incubated with ab316318 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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PYGO2 antibody [EPR27168-186] (AB316318)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling PYGO2 with ab316318 at 1/100 (5.13 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing mainly nuclear staining in HeLa cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PYGO2 antibody [EPR27168-186] (AB316318)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling PYGO2 with ab316318 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-PYGO2 antibody [EPR27168-186] (AB316318)

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-PYGO2 antibody [EPR27168-186] (ab316318) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

HeLa transfected with siRNA specifically targeting human PYGO2 whole cell lysate at 20 µg with 5% NFDM/TBST

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: 50 kDa,36 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-PYGO2 antibody [EPR27168-186] (AB316318)

In lane 5, the lysate was stored at -80? prior to Western Blotting. In lane 1, 2, 3 & 4, lysates were freshly made and used for Western Blotting immediately to minimize protein degradation

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

Exposure time : Lane 1-4 : 26 seconds; Lane 5 : 37 seconds

All lanes:

Western blot - Anti-PYGO2 antibody [EPR27168-186] (ab316318) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lanes 3 and 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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: 50 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-PYGO2 antibody [EPR27168-186] (AB316318)

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-PYGO2 antibody [EPR27168-186] (ab316318) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg with 5% NFDM/TBST

Lane 2:

Mouse testis tissue lysate at 20 µg with 5% NFDM/TBST

Lane 3:

Rat liver tissue lysate at 20 µg with 5% NFDM/TBST

Lane 4:

Rat testis tissue lysate at 20 µg with 5% NFDM/TBST

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: 50 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-PYGO2 antibody [EPR27168-186] (AB316318)

In Western blot, Anti-Myc tag mAb staining at 1/10000 dilution.

All lanes:

Western blot - Anti-PYGO2 antibody [EPR27168-186] (ab316318) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with human PYGO1 expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with 5% NFDM/TBST

Lane 2:

293T cells transfected with human PYGO2 expression vector containing a myc-His-tag®, whole cell lysate at 10 µg with 5% NFDM/TBST

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: 50 kDa

false

Exposure time: 10s