Anti-GPR177/WLS antibody [EPR29686-521]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPR177/WLS antibody [EPR29686-521] (AB323884)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling GPR177/WLS with ab323884 at 1/100 (5.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon carcinoma. The section was incubated with ab323884 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-GPR177/WLS antibody [EPR29686-521] (AB323884)

Immunohistochemical analysis of paraffin-embedded (A) HaCaT (Human skin keratinocyte) cell pellet, (B) A-204 (Human muscle rhabdomyosarcoma cell) cell pellet, (C) HepG2 (Human hepatocellular carcinoma epithelial cell) cell pellet, and (D) K562 (Human chronic myelogenous leukemia lymphoblast) cell pellet labeling GPR177/WLS with ab323884 at 1/1000 (0.501 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on HaCaT (image A) and A-204 (image B). No staining on HepG2 (image C) and K562 (image D). The section was incubated with ab323884 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-GPR177/WLS antibody [EPR29686-521] (AB323884)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling GPR177/WLS with ab323884 at 1/100 (5.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human kidney. The section was incubated with ab323884 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-GPR177/WLS antibody [EPR29686-521] (AB323884)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling GPR177/WLS with ab323884 at 1/100 (5.01 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast carcinoma. The section was incubated with ab323884 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-GPR177/WLS antibody [EPR29686-521] (AB323884)

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

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

All lanes:

Western blot - Anti-GPR177/WLS antibody [EPR29686-521] (ab323884) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 60 µg

Lane 2:

U-87 MG transfected with siRNA specifically targeting WLS whole cell lysate at 60 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: 45 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-GPR177/WLS antibody [EPR29686-521] (AB323884)

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

Negative control : HepG2, K-562.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-GPR177/WLS antibody [EPR29686-521] (ab323884) at 1/1000 dilution

Lane 1:

HaCaT (human skin keratinocyte) whole lysate at 60 µg

Lane 2:

A-204 (human muscle rhabdomyosarcoma cell) whole lysate at 60 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole lysate at 60 µg

Lane 4:

K-562 (human chronic myelogenous leukemia lymphoblast) whole lysate at 60 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: 45 kDa,124 kDa

false

Exposure time: 15s