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Immunofluorescence analysis of Smooth muscle actin was done on 70% confluent log phase MDCK cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with ABfinity™ Smooth muscle actin Recombinant Rabbit Monoclonal Antibody (701457) at 2 µg-4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (A12381). Panel d is a merged image showing actin filament localization of Smooth muscle actin. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 2–10 of human Smooth Muscle Actin|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||2-4µg/10^6 cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:500|
|Western Blot (WB)||1-3µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with bovine, canine, rabbit, rat and porcine based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Smooth muscle is found in the walls of blood vessels, lymphatic vessels, and a number of other tissues, where it provides mechanical support, and is a major component of the contractile apparatus. One of the primary components of smooth muscle is actin, which has six known isotypes in mammals. Smooth muscle actin exists in alpha-and gamma-actin isoforms.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
AAT6; actin; actin, alpha, vascular smooth muscle; Actin, aortic smooth muscle; ACTSA; alpha 2 actin; Alpha-actin-2; alpha-cardiac actin; alpha-sma; aortic smooth muscle; Cell growth-inhibiting gene 46 protein; GIG46; growth-inhibiting gene 46; MYMY5; smooth muscle alpha-actin
0610041G09Rik; a-SMA; AAT6; ACTA2; ACTSA; ACTVS; alphaSMA; GIG46; MYMY5; SMalphaA