Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

This data was developed using the same antibody clone in a different buffer formulation (ab7817).

Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling alpha smooth muscle actin with ab7817 at a concentration of 0.01 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab7817 anti-alpha smooth muscle actin antibody [1A4] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

This data was developed using the same antibody clone in a different buffer formulation (ab7817).

ab7817 staining ACTA2 in SaOS-2 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab7817 at 1µg/ml (shown in green) and Rabbit polyclonal to beta Tubulin - Loading Control (ab6046) (shown in pseudocolour magenta). Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

Immunohistochemical analysis of Human colon tissue labelling alpha smooth muscle Actin with ab7817 at 1/50000 dilution and ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) Secondary antibody. Positive cytoplasmic staining on Human colon tissue is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

This image is produced using the same clone but in a different formulation ab7817.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

Immunohistochemical analysis of Rat colon tissue labelling alpha smooth muscle Actin with ab7817 at 1/50000 dilution and ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) Secondary antibody. Positive cytoplasmic staining on Rat colon tissue is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

This image is produced using the same clone but in a different formulation ab7817.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

This data was developed using the same antibody clone in a different buffer formulation (ab7817).

ab7817 staining ACTA2 in NIH3T3 WT and NIH3T3-ACTA2 KO cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab7817 at 1µg/ml (shown in green) and Rabbit polyclonal to beta Tubulin - Loading Control (ab6046) (shown in pseudocolour magenta). Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

This data was developed using ab7817, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH 3T3 cells labelling alpha smooth muscle Actin with ab7817 at 1/50 dilution (Green). Confocal image showing cytoskeleton staining in NIH 3T3 cells. ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed was used as a secondary at 1/10 dilution. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/20 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/50 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Negative control : secondary counterstain antibody only control.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

This data was developed using ab7817, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH 3T3 cells labelling alpha smooth muscle Actin with ab7817 at 1/50 dilution (Green). Confocal image showing cytoskeleton staining in NIH 3T3 cells. ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed was used as a secondary at 1/10 dilution. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/20 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/50 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Negative control : secondary counterstain antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

Immunohistochemical analysis of Mouse colon tissue labelling alpha smooth muscle Actin with ab7817 at 1/50000 dilution and ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) Secondary antibody. Positive cytoplasmic staining on Mouse colon tissue is observed. Counterstained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody was ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

This image is produced using the same clone but in a different formulation ab7817.

• WB

Lab

Western blot - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

This image is produced using the same clone but in a different formulation ab7817.

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

All lanes:

Western blot - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (ab240654) at 1/1000 dilution

Lane 1:

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

Lane 2:

IMR-90 (human lung fibroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

false

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] - BSA and Azide free (AB240654)

This data was developed using the same antibody clone in a different buffer formulation (ab7817).

ab7817 staining alpha smooth muscle Actin in SV40LT-SMC cells (positive control top panel) and A431 cells (negative control bottom panel). The cells were fixed with 100% methanol (5 min) then 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 with ab7817 at 1μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).