Anti-alpha smooth muscle Actin 抗体 [1A4]

• IHC-P

Lab

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

AbReview60458****

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

ab7817 staining alpha smooth muscle Actin in human IMR-90 (Human Lung Fibroblast Cell Line) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde permeabilized with 0.1% TritonX-100 and blocked with 100% Cad-Block for 30 minutes at room temperature. Samples were incubated with primary antibody 3.41μg/ml in antibody diluent buffer for 16 hours at 4°C. An Alexa Fluor^® 488-conjugated polyclonal Goat anti-mouse IgG dilution 1/400 was used as secondary antibody.

This image is courtesy of an anonymous abreview.

• IHC-P

AbReview34173****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

Immunohistochemical analysis of mouse aorta (A) or skin (B) tissue staining alpha smooth muscle Actin with ab7817.

Tissue was fixed with 10% Neutral Buffered Formalin and blocked with 1% serum for 45 minutes 21°C; antigen retrieval was by enzymatic method in 0.0001% Trypsin-CaCl. Samples were incubated with primary antibody (0.034μg/ml in 0.3% Triton X-100 in PBS) for 1 hour at 21°C. A biotin-conjugated horse anti-mouse polyclonal IgG (1/50 dilution) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (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.

• ICC/IF

AbReview16561****

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

ab7817 staining alpha smooth muscle Actin in mouse heart cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde permeabilized with TritonX-100 and blocked with 5% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody 6.82μg/ml in blocking buffer for 2 hours. An Alexa Fluor^® 488-conjugated Donkey monoclonal to mouse IgG dilution 1/200 was used as secondary antibody.

This image is courtesy of an Abreview submitted by Dr. Ho-Jae Lee

• IHC-P

AbReview64448****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of 10% NBF-fixed human colon tissue permeabilized with 0.05% tween 20. Stained with ab7817 at 1/100 dilution. Secondary antibody used was Alexa fluor® 488 Donkey anti-Rabbit IgG at 1/300 dilution. Blocking was done with Sea Block for 30 minutes at 22°C. The sample was incubated with the primary antibody and Sea Block for 14 hours at 4°C. Antigen retrieval method was heat mediated, ab94674 100X Citrate Buffer pH 6.0.

This image is courtesy of an Abreview submitted by David Ivancic

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

Overlay histogram showing SV40LT-SMC cells stained with ab7817 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab7817 1.137μg/ml) for 30 min at 22°C. The secondary antibody used was Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) (ab150113) at 1/2000 dilution for 30 min at 22°C

Isotype control antibody (black line) was mouse IgG2a [18C8BC7AD10] (ab170191) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (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.

• IHC-P

AbReview60459****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

ab7817 staining alpha smooth muscle actin in Mouse intestine tissue by Immunohistochemistry-Immunofluorescence. Tissue was fixed with formaldehyde and blocked with 100% Cas-block for 30 minutes at room temperature; antigen retrieval was performed by heat mediated citrate buffer, pH6. The sample was incubated with primary antibody at 0.034μg/ml for 16 hours at 4°C. An Alexa Fluor^® 488 Goat anti-mouse IgG was used as the secondary antibody at 1/400 dilution. Autofluorescence was blocked with 0.1% Sudan Black in 70% ethanol for 10 minutes at room temperature after antigen retrieval, and followed with 3X wash with PBS-T after antigen retrieval. Image was taken with confocal microscope.

This image is courtesy of an anonymous Abreview

• IHC-P

AbReview39194****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

ab7817 staining alpha smooth muscle actin in Human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde and permeabilized with wash buffer with tween; antigen retrieval was by heat mediation in Tris-EDTA buffer pH 9.0. Samples were incubated with primary antibody (0.034μg/ml in blocking buffer) for 30 minutes at 20°C. A HRP-conjugated Goat anti-mouse IgG polyclonal (undiluted) was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Rudolf Jung

• ICC/IF

AbReview36904****

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

ab7817 staining alpha smooth muscle Actin (green) in Mouse primary colon myofibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone and blocked with 5% BSA for 30 hours at 25°C. Samples were incubated with primary antibody (1/100 dilution in PBS + 5% BSA) for 2 hours at 25°C. Donkey Anti-Mouse IgG H&L (DyLight^® 488) (ab96875) (1/1000 dilution) was used as the secondary antibody. Costained with ab92547 Rabbit anti-Vimentin (red).

This image is courtesy of an Abreview submitted by Charles Pallangyo

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [1A4] (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).

• WB

Lab

Western blot - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

Gel type : MOPS

Blocking buffer : 3% milk

Loading control : alpha tubulin (ab52866), secondary Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (1 : 10000 dilution)

All lanes:

Western blot - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1 µg/mL

Lane 1:

NIH 3T3 whole cell lysate at 20 µg

Lane 2:

SV40LT-SMC whole cell lysate at 20 µg

Lane 3:

A431 whole cell lysate at 20 µg

Lane 4:

A549 whole cell lysate at 20 µg

Lane 5:

Jurkat whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L (IRDye® 800RD) at 1/10000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

• WB

Lab

Western blot - Anti-alpha smooth muscle Actin antibody [1A4] (AB7817)

Gel type : MOPS

Blocking buffer : 3% milk

Loading control : alpha tubulin (ab52866), secondary Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (1 : 10000 dilution)

All lanes:

Western blot - Anti-alpha smooth muscle Actin antibody [1A4] (ab7817) at 1 µg/mL

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Human Foreskin Fibroblast Whole Cell Lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L (IRDye® 800RD) at 1/10000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false