All lanes : Anti-gamma Catenin antibody [EPR17310] (ab184919) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : JUZ knockout HEK-293T cell lysate
Lane 3 : A431 cell lysate
Lane 4 : THP-1 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 82 kDa
Observed band size: 82 kDa

Lanes 1-4: Merged signal (red and green). Green - ab184919 observed at 82 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab184919 Anti-gamma Catenin antibody [EPR17310] was shown to specifically react with gamma Catenin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266272 (knockout cell lysate ab257269) was used. Wild-type and gamma Catenin knockout samples were subjected to SDS-PAGE. ab184919 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

 

All lanes : Anti-gamma Catenin antibody [EPR17310] (ab184919) at 1/1000 dilution

Lane 1 : Human fetal heart lysates
Lane 2 : Human fetal kidney lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 82 kDa
Observed band size: 82 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time = 15 seconds

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) labelling gamma Catenin with purified ab184919 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling gamma Catenin with ab184919 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and membrane staining on epithelial cells of Human prostate is observed; and smooth muscle cells are also positively stained. Counterstained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling gamma Catenin with ab184919 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and membrane staining on epithelial cells of Human skin is observed. Counterstained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling gamma Catenin with ab184919 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasmic and membrane staining on epithelial cells of mouse stomach is observed. Counterstained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded rat skin tissue labeling gamma Catenin with ab184919 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Membrane and cytoplasmic staining on the epithelial cells of rat skin is observed; and smooth muscle cells are also positively stained. Counterstained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling gamma Catenin with ab184919 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab184919 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDCK (Canine kidney cell line) cells labeling gamma Catenin with ab184919 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing membranous staining on MDCK cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab184919 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

 

All lanes : Anti-gamma Catenin antibody [EPR17310] (ab184919) at 1/10000 dilution

Lane 1 : Human fetal skin lysates
Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 82 kDa
Observed band size: 82 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time = 3 seconds

All lanes : Anti-gamma Catenin antibody [EPR17310] (ab184919) at 1/1000 dilution

Lane 1 : Hela (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : Mouse skin lysates
Lane 3 : Rat skin lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 82 kDa
Observed band size: 82 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time = 5 seconds

Anti-gamma Catenin antibody [EPR17310] (ab184919) at 1/1000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg

Secondary
Mouse monoclonal to cardiac Troponin I (ab1000) (Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated)

Predicted band size: 82 kDa
Observed band size: 82 kDa

Blocking and diluting buffer and concentration = 5% NFDM/TBST

Exposure time = 3 minutes

Anti-gamma Catenin antibody [EPR17310] (ab184919) at 1/100000 dilution + MDCK (Canine kidney cell line) whole cell lysate at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 82 kDa
Observed band size: 82 kDa


Exposure time: 1 minute

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