Alpha-Smooth Muscle Actin Recombinant Rabbit Monoclonal Antibody (17H19L35)

货号：701457

规格：100 µg

价格：4885

产品类型：免疫组化一抗

品牌：eBioscience

物种：人

宿主：兔

抗体亚型：IgG

荧光染料：Purified

类型:	单抗	同型对照：
浓度：	0.5 mg/mL	用法：	2-4 µg(ICC);2-4 µg(IF);1:50-1:500（IHC (P)）；1-3 µg/mL(WB)
免疫原性：	Peptide corresponding to amino acids 2–10 of human Smooth Muscle Actin

产品详细信息This antibody is predicted to react with bovine, canine, rabbit, rat and porcine based on sequence homology.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.Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.靶标信息Smooth Muscle Actin belongs to the actin family of proteins, which are highly conserved proteins that play a role in cell motility, structure and integrity. Alpha, beta and gamma actin isoforms have been identified, with alpha actin being a major constituent of the contractile apparatus, while beta and gamma actins are involved in the regulation of cell motility. In particular, smooth muscle actin is an alpha actin that is found in skeletal muscle. Actin exists as a ubiquitous protein involved with filament formation that make up large portions of the cytoskeleton. Actin filaments interact with myosin to assist in muscle contraction as well as aiding in cell motility and cytokinesis. Smooth muscle actin is found on smooth muscle vessel walls, gut wall, myometrium, myoepithelial cells in breast and salivary glands. Defects in the smooth muscle actin gene cause aortic aneurysm familial thoracic type 6. Actin isoforms differ slightly in their N-terminus and the sequences of each are perfectly conserved in higher vertebrates. Alpha-smooth muscle actin is abundant in vascular and visceral smooth muscle cells. In addition, it has also been shown that smooth muscle actin appear in stress fibers of fibroblastic cells during pathological situations involving contractile phenomena such as wound healing and fibrocontractive diseases. Multiple alternatively spliced variants of smooth muscle actin have been identified.

数据

Alpha-Smooth Muscle Actin Antibody (701457) in IHC (P)Immunohistochemistry analysis of Smooth Muscle Actin showing staining in the cytoplasm of paraffin-embedded human uterus tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with Smooth Muscle Actin monoclonal antibody (Product # 701457) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using a HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting. Alpha-Smooth Muscle Actin Antibody (701457) in IFImmunofluorescent 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 Smooth muscle actin Recombinant Rabbit Monoclonal Antibody (Product # 701457) at 2 µg-4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) 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 (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # 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. Alpha-Smooth Muscle Actin Antibody (701457) in WBWestern blot analysis of Smooth Muscle Actin in whole cell extracts from rat heart using a Smooth Muscle Actin recombinant rabbit monoclonal antibody (Product # 701457) at a dilution of 1 µg/mL. Detection was performed using an HRP-conjugated goat anti-rabbit secondary antibody followed by chemiluminescence (ECL). Results show a band at ~40kDa. Alpha-Smooth Muscle Actin Antibody (701457) in WBWestern blot analysis of Smooth Muscle Actin was performed by loading 20 µg of Rat Heart (lane1) and Mouse Heart (lane2) tissue lysates using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. Smooth Muscle Actin was detected at ~40 kDa using Smooth Muscle Actin Recombinant Rabbit Monoclonal Antibody (Product # 701457) at 1 µg-3 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat anti-Rabbit IgG-HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

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参考文献：

1.Biology of sex differencesSevere pulmonary hypertension in aging female apolipoprotein E-deficient mice is rescued by estrogen replacement therapy. 2.Atherosclerosis CD98 regulates vascular smooth muscle cell proliferation in atherosclerosis. "701457 was used in immunohistochemistry to ask if CD98 heavy chain expressed on vascular smooth muscle cells plays a role in shaping the morphology of atherosclerotic plaques by regulating vascular smooth muscle cell function" 3.Biology of sex differencesSevere pulmonary hypertension in aging female apolipoprotein E-deficient mice is rescued by estrogen replacement therapy.

技术参数

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