Anti-gamma Catenin antibody [11E4] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma Catenin antibody [11E4] - BSA and Azide free (AB269564)

IHC image of gamma Catenin staining in a section of formalin-fixed paraffin-embedded normal human cervix* performed on a Leica BOND^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab231304, 1ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

This image was produced using the same antibody clone but in a different formulation ab231304, PBS and sodium azide.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-gamma Catenin antibody [11E4] - BSA and Azide free (AB269564)

ab231304 staining gamma Catenin in A431 cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1%PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab231304 at 1μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This image was produced using the same antibody clone but in a different formulation ab231304, PBS and sodium azide.

• WB

Lab

Western blot - Anti-gamma Catenin antibody [11E4] - BSA and Azide free (AB269564)

This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab231304 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

This image was produced using the same antibody clone but in a different formulation ab231304, PBS and sodium azide.

All lanes:

Western blot - Anti-gamma Catenin antibody [11E4] (<a href='/en-us/products/primary-antibodies/gamma-catenin-antibody-11e4-ab231304'>ab231304</a>) at 1 µg/mL

Lane 1:

A431 whole cell lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Predicted band size: 82 kDa

Observed band size: 25 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-gamma Catenin antibody [11E4] - BSA and Azide free (AB269564)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before ab231304 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/10000 dilution respectively. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-gamma Catenin antibody [11E4] (<a href='/en-us/products/primary-antibodies/gamma-catenin-antibody-11e4-ab231304'>ab231304</a>)

Lane 1:

A431 whole cell lysate at 20 µg

Lane 2:

HeLa whole cell lysate at 20 µg

Predicted band size: 82 kDa

Observed band size: 90 kDa

false