Anti-beta Catenin antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody (AB6302)

This picture was kindly supplied as part of the review submitted by Mohaiza Dashwood. Immunofluorescence of H29 cells stained with DAPI (blue) and rabbit polyclonal anti-beta-catenin (ab6302), 1/2000) with Alexa Fluor^® 488 (green) from Molecular Probes.

• ICC/IF

AbReview13479****

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody (AB6302)

ab6302 at 1/500 dilution staining beta Catenin in human breast cells by immunocytochemistry/ immunofluorescence. Sections were formaldehyde fixed, permeabilized in 0.5% digitonin prior to incubating with ab6302 for 1 hour. Alexa fluor^® 488 goat polyclonal, diluted 1/500, was used as the secondary antibody. Treated samples received 10um GSK-3 Inhibitor X.

This image is courtesy of an Abreview submitted by Roderick Benson

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody (AB6302)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling beta Catenin with ab320753 at 1/10000 (0.05 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver. The section was incubated with ab320753 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Recombinant multiclonal ab320753 at 1/10, 000 (left) showed similar staining pattern to the polyclonal ab6302 at 1/10, 000 (right).

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

AbReview15665****

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody (AB6302)

ab6302 at 1/2000 dilution staining beta Catenin in human HeLa cells by immunocytochemistry/ immunofluorescence. Sections were paraformaldehyde fixed, permeabilized in 0.3% Triton X prior to blocking in 5% BSA for 2 hours at 27°C and then incubated with ab6302 for 8 hour at 4°C. Alexa fluor^® 488 goat polyclonal, diluted 1/1000, was used as the secondary antibody. Counterstaining with DAPI.

This image is courtesy of an Abreview submitted by Jennifer Schnabel

• IP

Supplier Data

Immunoprecipitation - Anti-beta Catenin antibody (AB6302)

beta Catenin was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab320753 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320753 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab320753 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab320753 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

Exposure time : Lanes 1-3 : 3 seconds, Lanes 4-6 : 6 seconds

All lanes:

Immunoprecipitation - Anti-beta Catenin antibody [RM1213] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-rm1213-ab320753'>ab320753</a>) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

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

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• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody (AB6302)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling beta Catenin with ab6302 at 1/20,000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody (AB6302)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling beta Catenin with ab6302 at 1/20,000 dilution.

• WB

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Western blot - Anti-beta Catenin antibody (AB6302)

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

In Western blot, ab320753 and ab6302 were shown to bind specifically to CTNNB1. Target of interest was observed at 85 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in CTNNB1 knockout cell line (lane 2).

Recombinant multiclonal ab320753 (left) showed higher sensitivity than Polyclonal ab6302 (right).

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

Exposure time : Lane 1-2 : 103 seconds Lane 3-6 : 180 seconds

All lanes:

Western blot - Anti-beta Catenin antibody [RM1213] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-rm1213-ab320753'>ab320753</a>) at 1/20000 dilution

Lane 1:

Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

CTNNB1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

NIH/3T3 (mouse embryonic fibroblast) whole cell 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 1/100000 dilution

Observed band size: 85 kDa,36 kDa

false

• WB

AbReview4458****

Western blot - Anti-beta Catenin antibody (AB6302)

This image is courtesy of an Abreview submitted by Mike Campa on 4 April 2006.

All lanes:

Western blot - Anti-beta Catenin antibody (ab6302) at 1/4000 dilution

Lane 1:

5ug human lung tumour lysate.

Lane 2:

10ug human lung tumour lysate.

Lane 3:

20ug human lung tumour lysate.

Secondary

All lanes:

Goat anti-Rabbit IgG (H&L)HRP

Predicted band size: 85 kDa

Observed band size: 95 kDa

false

Exposure time: 10s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody (AB6302)

Immunocytochemistry analysis of bovine kidney cells labeling beta Catenin with ab6302 at 1/20,000 dilution. Cells were fixed and permeabilized with Methanol followed by Acetone. The antibody was developed using Anti-Rabbit IgG (whole molecule)-FITC antibody produced in Goat. Cells were counterstained with DAPI (blue) to stain nuclei.

• WB

Supplier Data

Western blot - Anti-beta Catenin antibody (AB6302)

All lanes:

Western blot - Anti-beta Catenin antibody (ab6302) at 1/8000 dilution

All lanes:

Bovine kidney tissue lysate

Predicted band size: 85 kDa

false