Anti-beta Catenin antibody [EP690Y] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-beta Catenin antibody [EP690Y] - BSA and Azide free (AB247372)

Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling beta Catenin with Purified ab68183 at 1/20 dilution (5 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68183).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [EP690Y] - BSA and Azide free (AB247372)

Immunocytochemistry/ Immunofluorescence analysis of parental HAP1 (Wildtype control Human chronic myelogenous leukemia near-haploid cell line) cells labeling beta Catenin with Purified ab68183 at 1/100 dilution (10 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68183).

• WB

Lab

Western blot - Anti-beta Catenin antibody [EP690Y] - BSA and Azide free (AB247372)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68183). Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab181602 was used as a GAPDH loading control. Raw264.7 expresses low level of beta Catenin and stimulation is required to allow detection of the beta Catenin protein in this cell line, as described in PMID : 22983902 and PMID : 29137395.

All lanes:

Western blot - Anti-beta Catenin antibody [EP690Y] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-ep690y-ab68183'>ab68183</a>) at 1/1000 dilution

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell) 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 1/20000 dilution

Predicted band size: 86 kDa

Observed band size: 90 kDa

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Exposure time: 180s

• WB

Lab

Western blot - Anti-beta Catenin antibody [EP690Y] - BSA and Azide free (AB247372)

False colour image of Western blot : Anti-beta Catenin antibody [EP690Y] staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab68183 was shown to bind specifically to beta Catenin. A band was observed at 85 kDa in wild-type HepG2 cell lysates with no signal observed at this size in CTNNB1 knockout cell line. To generate this image, wild-type and CTNNB1 knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-beta Catenin antibody [EP690Y] - BSA and Azide free (ab247372) at 1/500 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

CTNNB1 knockout HepG2 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

A431 cell lysate at 20 µg

Predicted band size: 85 kDa

Observed band size: 85 kDa

false

• WB

Unknown

Western blot - Anti-beta Catenin antibody [EP690Y] - BSA and Azide free (AB247372)

full-length beta catenin : 90kDa ; C-terminal cleavage fragment : 75kDa (PMID : 15492240).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68183).

All lanes:

Western blot - Anti-beta Catenin antibody [EP690Y] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-ep690y-ab68183'>ab68183</a>) at 1/1000 dilution

All lanes:

A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 15 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

Predicted band size: 85 kDa

Observed band size: 75 kDa,90 kDa

false

• WB

Unknown

Western blot - Anti-beta Catenin antibody [EP690Y] - BSA and Azide free (AB247372)

full-length beta catenin : 90kDa ; C-terminal cleavage fragment : 75kDa (PMID : 15492240).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68183).

All lanes:

Western blot - Anti-beta Catenin antibody [EP690Y] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-ep690y-ab68183'>ab68183</a>) at 1/1000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg

Lane 2:

C6 (Rat glial tumor glial cell) whole cell lysates 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

Predicted band size: 85 kDa

Observed band size: 75 kDa,90 kDa

false