重组Anti-beta Catenin (phospho S675)抗体[EPR28410-45]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling beta Catenin (phospho S675) with ab314450 at 1/1000 (0.497 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Membranous staining on (A) human colon without alkaline phosphatase treatment; no staining on (B) human colon with alkaline phosphatase treatment. The section was incubated with ab314450 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling beta Catenin (phospho S675) with ab314450 at 1/1000 (0.497 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Membranous staining on (A) human breast carcinoma without alkaline phosphatase treatment; no staining on (B) human breast carcinoma with alkaline phosphatase treatment. The section was incubated with ab314450 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

beta Catenin (phospho S675) was immunoprecipitated from 0.35 mg SK-N-MC (human brain epithelial cell) treated with 100 uM forskolin for 30 minutes whole cell lysate with ab314450 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314450 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (ab314450) at 1/1000 dilution

Lane 1:

SK-N-MC (human brain epithelial cell) treated with 100 μM forskolin for 30 minutes whole cell lysate

Lane 2:

ab314450 at 1/30 IP in SK-N-MC (human brain epithelial cell) treated with 100 uM forskolin for 30 minutes whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab314450 in SK-N-MC treated with 100 uM forskolin for 30 minutes whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse colon (fresh frozen) tissue labeling beta Catenin (phospho S675) with ab314450 at 1/500 (0.994 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on mouse colon, while nearly no staining on mouse colon after phosphatase treatment. The nuclear counterstain was DAPI (Blue). The section was incubated with ab314450 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling beta Catenin (phospho S675) with ab314450 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Membranous staining on (A) mouse breast cancer without alkaline phosphatase treatment; no staining on (B) mouse breast cancer with alkaline phosphatase treatment. The section was incubated with ab314450 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling beta Catenin (phospho S675) with ab314450 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Membranous staining on (A) mouse colon without alkaline phosphatase treatment; no staining on (B) mouse colon with alkaline phosphatase treatment. The section was incubated with ab314450 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling beta Catenin (phospho S675) with ab314450 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Membranous staining on (A) rat colon without alkaline phosphatase treatment; no staining on (B) rat colon with alkaline phosphatase treatment. The section was incubated with ab314450 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

beta Catenin (phospho S675) was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab314450 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314450 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (ab314450) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate

Lane 2:

ab314450 at 1/30 IP in Mouse brain tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab314450 in mouse brain tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• WB

Supplier Data

Western blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 30806153).

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

In Western blot, Anti-beta Catenin antibody - Total protein control (ab32572) staining at 1/1000 dilution.

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

All lanes:

Western blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (ab314450) at 1/1000 dilution

Lane 1:

Untreated SK-N-MC (human brain epithelial cell) whole cell lysate (untreated membrane) at 40 µg

Lane 2:

SK-N-MC treated with 100 uM forskolin for 30 minutes whole cell lysate (untreated membrane) at 40 µg

Lane 3:

Untreated SK-N-MC (human brain epithelial cell) whole cell lysate (Alkaline phosphatase treated membrane) at 40 µg

Lane 4:

SK-N-MC treated with 100 uM forskolin for 30 minutes whole cell lysate (Alkaline phosphatase treated membrane) at 40 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: 85 kDa,90 kDa,36 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Dot blot analysis of beta Catenin (phospho S675) using ab314450 at 1 : 1000 (0.497 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 1000 (0.497 ug/ml) dilution.

Exposure time : 180 Seconds.

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

This antibody does not cross-react with gamma Catenin (phospho S665).

• WB

Supplier Data

Western blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

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

In Western blot, Anti-beta Catenin antibody - Total protein control (ab32572) staining at 1/1000 dilution.

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

All lanes:

Western blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (ab314450) at 1/1000 dilution

Lane 1:

Mouse colon tissue lysate (untreated membrane) at 20 µg

Lane 2:

Rat liver tissue lysate (untreated membrane) at 20 µg

Lane 3:

Mouse colon tissue lysate (Alkaline phosphatase treated membrane) at 20 µg

Lane 4:

Rat liver tissue lysate (Alkaline phosphatase treated membrane) 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: 85 kDa,90 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

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

The identity of the lower MW band at approximately 37 kDa in Lane 2 is unknown.

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

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

All lanes:

Western blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (ab314450) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse testis tissue 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: 90 kDa,36 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-beta Catenin (phospho S675) antibody [EPR28410-45] (AB314450)

Dot blot analysis of beta Catenin (phospho S675) using ab314450 at 1 : 1000 (0.497 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 1000 (0.497 ug/ml) dilution.

Exposure time : 180 Seconds.

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