Recombinant Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] - BSA and Azide free (ab215368)

FoxA1 (red) and alpha smooth muscle actin (green) staining are shown by indirect immunofluorescence on sections of prostate from mice of the indicated genotypes.

Wild type: 21 weeks, Tgfbr2^r/r: 44 weeks, Pten^r/r: 21 weeks, Pten^r/r;Tgfbr2^r/r: 11 weeks, Apc^r/r: 36 weeks, and Apc^r/r;Tgfbr2^r/r: 24 weeks old.

IF images were captured on an Olympus BX51 microscope and DP70 digital camera, or on a Nikon Eclipse NI-U and captured with a DS-QI1 camera with NIS Elements software.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32575).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labeling alpha smooth muscle Actin (green) with purified ab32575 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 Alexa Fluor^®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).

For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).  

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32575).

Intracellular Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling  alpha smooth muscle Actin with purified ab32575 at a dilution of 1/20 (red). 

Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies. 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32575). 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse smooth muscle tissue labeling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody.

Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32575).

This IHC data was generated using the same anti-alpha smooth muscle Actin antibody clone, E184, in a different buffer formulation (cat# ab32575).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human smooth muscle tissue labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

Clone E184 (ab215368) has been successfully conjugated by Abcam. This image was generated using Anti-alpha smooth muscle Actin antibody [E184] (Alexa Fluor® 488). Please refer to ab197240 for protocol details.

ab197240 staining alpha smooth muscle actin in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 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 ab197240 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).

Clone E184 (ab215368) has been successfully conjugated by Abcam. This image was generated using Anti-alpha smooth muscle Actin antibody [E184] (PE). Please refer to ab209435 for protocol details.

ab209435 staining alpha smooth muscle Actin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 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 ab209435 at 1/500 dilution (Pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

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

Clone E184 (ab215368) has been successfully conjugated by Abcam. This image was generated using Anti-alpha smooth muscle Actin antibody [E184] (Alexa Fluor® 647). Please refer to ab196919 for protocol details.

ab196919 staining alpha Smooth Muscle Actin in HeLa cells. The cells were fixed with 4% PFA (10min), permeabilized with 0.1% Triton X-100 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 ab196919 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 2µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

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

Following reconstitution in PBS with 10% DMSO, peptides (1 ug per mL) were immobilised in PBS on an ELISA plate overnight. After blocking in 5% BSA, primary antibody (ab271180) was added in a concentration range of 0.017-1000 ng per mL.

Pre-adsorbed secondary antibody goat anti-rabbit IgG H&L (HRP, ab97080) was used at 1/20000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32575).

Following reconstitution in PBS with 10% DMSO, peptides (1 ug per mL) were immobilised in PBS on an ELISA plate overnight. After blocking in 5% BSA, primary antibody (ab271180) was added in a concentration range of 0.017-1000 ng per mL.

Pre-adsorbed secondary antibody goat anti-rabbit IgG H&L (HRP, ab97080) was used at 1/20000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32575).