Alpha-Smooth Muscle Actin Monoclonal Antibody (1A4 (asm-1))

货号：MA5-11547

规格：500 µL

价格：4885

产品类型：免疫组化一抗

品牌：eBioscience

物种：人/小鼠/大鼠

宿主：小鼠

抗体亚型：IgG2a, kappa

荧光染料：Purified

类型:	单抗	同型对照：
浓度：	40 µg/mL	用法：	1:100-1:500（ICC）；1:100-1:500（IF）；1:800（IHC (P)）；1:100-1:500（WB）
免疫原性：	N-terminal decapeptide of alpha-smooth muscle isoform of actin; acetylated at the N-terminus

产品详细信息MA5-11547 targets Actin Smooth Muscle IF, WB, and IHC (P) applications and shows reactivity with Bovine, Chicken, Human, mouse, Non-human primate, Rabbit, and Rat samples. This antibody detects a non-specific band at approx. 30 kDa in Hela cell lysates.The MA5-11547 immunogen is n-terminal decapeptide of alpha-smooth muscle isoform of actin; acetylated at the N-terminus.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 (MA5-11547) in IHCFormalin-fixed, paraffin-embedded human leiomyoma stained with Smooth Muscle Actin antibody using peroxidase-conjugate and DAB chromogen. Note cytoplasmic staining of smooth muscle cells. Alpha-Smooth Muscle Actin Antibody (MA5-11547) in IFImmunofluorescent analysis of Actin Smooth Muscle (green) showing positive staining in the cytoplasm of Hela cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with an Actin Smooth Muscle monoclonal antibody (Product # MA5-11547) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x. Alpha-Smooth Muscle Actin Antibody (MA5-11547) in IFImmunofluorescent analysis of Actin Smooth Muscle (green) showing positive staining in the cytoplasm of C2C12 cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with an Actin Smooth Muscle monoclonal antibody (Product # MA5-11547) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x. Alpha-Smooth Muscle Actin Antibody (MA5-11547) in IFImmunofluorescent analysis of Actin Smooth Muscle (green) showing positive staining in the cytoplasm of A431 cells (right) compared with a negative control in the absence of primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes, blocked with 3% BSA-PBS for 30 minutes at room temperature and probed with an Actin Smooth Muscle monoclonal antibody (Product # MA5-11547) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 488-conjugated goat-anti-mouse IgG (H+L) secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with DAPI for 5-10 minutes in the dark. Images were taken at a magnification of 60x. Alpha-Smooth Muscle Actin Antibody (MA5-11547) in WBWestern blot analysis of Actin Sooth Muscle was performed by loading 25 µg of C2C12 (Lane 1), Hela (Lane 2), and A431 (Lane 3) cell lysates and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4°C overnight. The membrane was probed with an Actin Sooth Muscle monoclonal antibody (Product # MA5-11547) at a dilution of 1:100 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Results show a band at 42 kDa in all three cell lysates. Alpha-Smooth Muscle Actin Antibody (MA5-11547) in IFImmunofluorescence analysis of mesoderm using anti-smooth muscle actin antibody (Product # MA5-11547). Embryoid bodies (EBs) were generated from Gibco ® Human Episomal iPSC Line grown on Geltrex® in Essential 8TM Medium. After 3 weeks in culture, EBs were dissociated with TrypLETM and re-plated onto Geltrex®-coated multi-well plates. Cells were fixed, permeabilized and blocked for immunostaining. Cells were stained with a smooth muscle actin monoclonal antibody (Product # MA5-11547) at a dilution of 1:100 in 3% BSA/PBS blocking buffer overnight at 4°C, and then incubated with Alexa Fluor® 488 donkey anti mouse IgG2a antibody (Product # A-21131, green) at 1:500 dilution in conjunction with NucBlue® Fixed Cell Ready Probes® Reagent. After another 3 washes, the cells were imaged on EVOS®Floid® Cell Imaging system.

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参考文献：

1.Journal of visualized experiments : JoVEScaling of Engineered Vascular Grafts Using 3D Printed Guides and the Ring Stacking Method.2.Nature protocolsIsolation, culture and evaluation of multilineage-differentiating stress-enduring (Muse) cells.3.Cancer investigationStromal-epithelial crosstalk provides a suitable microenvironment for the progression of ovarian cancer cells in vitro.4.Cancer lettersMammary fibroblasts regulate morphogenesis of normal and tumorigenic breast epithelial cells by mechanical and paracrine signals.5.Cardiovascular researchHigh-purity enrichment of functional cardiovascular cells from human iPS cells.

技术参数

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