The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.3 - 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 42 kDa).
Use at an assay dependent concentration.
Antibody detection limit dilution 1:4,000.
Use a concentration of 2 - 4 µg/ml.
Use at an assay dependent concentration. PubMed: 20947685
Permeabilisation with ~0.2% Triton (in PBS) is recommended
Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
Involvement in disease
Defects in ACTA2 are the cause of aortic aneurysm familial thoracic type 6 (AAT6) [MIM:611788]. AATs are characterized by permanent dilation of the thoracic aorta usually due to degenerative changes in the aortic wall. They are primarily associated with a characteristic histologic appearance known as 'medial necrosis' or 'Erdheim cystic medial necrosis' in which there is degeneration and fragmentation of elastic fibers, loss of smooth muscle cells, and an accumulation of basophilic ground substance.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab21027)Image courtesy of an anonymous Abreview.
ab21027 staining alpha smooth muscle Actin in mouse pancreas by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 1% BSA for 3 hours at RT; antigen retrieval was by heat mediation in a citrate buffer pH6. Samples were incubated with primary antibody (1/100) for 16 hours. An Alexa Fluor® 647 donkey anti-goat IgG polyclonal (1/500) was used as the secondary antibody..
Western blot - Anti-alpha smooth muscle Actin antibody (ab21027)
All lanes : Anti-alpha smooth muscle Actin antibody (ab21027) at 0.1 µg/ml
Lane 1 : Human Duodenum tissue lysates Lane 2 : Heart tissue lysates
Lysates/proteins at 35 µg per lane.
Predicted band size: 42 kDa
Primary incubation was 1 hour. Detected by chemiluminescence.
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-alpha smooth muscle Actin antibody (ab21027)Image courtesy of an anonymous Abreview.
ab21027 staining alpha smooth muscle Actin in mouse femoral artery by Immunohistochemistry (PFA perfusion fixed frozen sections.) Tissue was fixed in formaldehyde, permeabilized using 0.1% Triton, blocked using 10% serum for 1 hour at 22°C and then incubated with ab21027 at a 1/300 dilution for 16 hours at 4 °C. The secondary used was an Alexa Fluor®488 rabbit anti-goat used at a 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human (top) and mouse (bottom) lung tissue sections labelling alpha smooth muscle Actin with ab21027 at 2.5µg/ml. Image shows staining of the alveolar walls. Trypsin-mediated antigen retrieval, HRP-staining. Bar in right lower corner represents 50um.