重组Anti-alpha smooth muscle Actin抗体[SP171] - BSA and Azide free

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Fluorescence multiplex immunohistochemical analysis of human liver tissue (formalin-fixed paraffin-embedded section). Panel A shows merged staining of ab195872 anti-Cytokeratin 19 stained on branch of bile ducts (magenta; Opal™690) at 1 : 8000 ( 0.127 μg/ml) [Panel B], ab275376 anti-Factor VIII stained on endothelial cells (red; Opal™570) at 1 : 1000 ( 0.457 μg/ml) [Panel C], and ab150301 anti-alpha smooth muscle Actin stained on smooth muscles (green; Opal™520) at 1 : 200 ( 0.14 μg/ml) [Panel C] on human liver. DAPI was used as a nuclear counter stain. Followed by Opal Polymer HRP Ms + Rb secondary. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. The section was incubated in three rounds of staining : in the order of ab195872 for 30 mins, ab275376 for 30 mins and ab150301 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins was used.  This data was developed using ab150301, the same antibody clone in a different buffer formulation.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Immunohistochemical analysis of formalin fixed, paraffin embedded human colon tissue labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab150301)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/200 dilution (0.825μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

This data was developed using the same antibody clone in a different buffer formulation (ab150301). ab150301 staining alpha smooth muscle Actin in wild-type HeLa cells (top panel) and ACTA2 knockout HeLa cells (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 ab150301 at 5μg/ml concentration and ab7291 (Mouse monoclonal to alpha 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 rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (ab150120) 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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301)

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat colon tissue sections labeling alpha smooth muscle Actin with ab150301 at 1/200 dilution (0.26 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10 mins. Goat Anti-Rabbit & Mouse IgG (HRP) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301)

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Intracellular Flow Cytometry analysis of C6 (Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/200 dilution (0.825μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/100(1.65 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab150301 at 1/200 dilution (0.825μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab150301).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody [SP171] - BSA and Azide free (AB242395)

This data was developed using the same antibody clone in a different buffer formulation (ab150301). ab150301 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 ab150301 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha 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 rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (ab150120) 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).