alpha-Smooth Muscle Actin Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-11075
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for alpha-Smooth Muscle Actin Overexpression Lysate
alpha-Smooth Muscle Actin Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC227802
Accession#: NM_001141945
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Expression Host: HEK293T
Plasmid: RC227802
Accession#: NM_001141945
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Product Specifications
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
TMW
42 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Type
Overexpression
Scientific Data Images for alpha-Smooth Muscle Actin Overexpression Lysate
![Western Blot: alpha-Smooth Muscle Actin Overexpression Lysate [NBP2-11075]](https://resources.bio-techne.com/images/products/Actin-Alpha-2-Smooth-Muscle-Overexpression-Lysate-Adult-Normal-Western-Blot-NBP2-11075-img0001.jpg "Western Blot: alpha-Smooth Muscle Actin Overexpression Lysate [NBP2-11075]")
Western Blot: alpha-Smooth Muscle Actin Overexpression Lysate [NBP2-11075]
Western Blot: alpha-Smooth Muscle Actin Overexpression Lysate [NBP2-11075] - (Left) - Empty vector transfected control cell lysate (HEK293 cell lysate); (Right) -Over-expression Lysate for alpha-Smooth Muscle Actin. An observed molecular weight of ~45 kDa is indicated here.Formulation, Preparation, and Storage
Formulation
RIPA buffer
Concentration
The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store at -80C. Avoid freeze-thaw cycles.
Background: alpha-Smooth Muscle Actin
ACTA2 encodes alpha Smooth Muscle Actin and is also known as alpha-actin, alpha-actin-2, aortic smooth muscle or alpha smooth muscle actin (alpha-SMA, SMactin, alpha-SM-actin, ASMA). Alpha Smooth Muscle Actin is frequently used as a marker of smooth muscle differentiation and has a theoretical molecular weight of 42 kDa. Smooth muscle alpha actin is one of a few genes whose expression is predominantly expressed in vascular smooth muscle cells that is used to study myofibroblasts and fibrosis (1). Excessive accumulation of alpha smooth muscle actin-positive (ACTA2+) activated myofibroblasts is observed in Idiopathic pulmonary fibrosis (2). Expression of smooth muscle alpha actin is regulated by cell proliferation and is altered in pathological conditions including atherosclerosis and is common in metastatic cancers (1,3).
References
1. Chakraborty, R., Saddouk, F. Z., Carrao, A. C., Krause, D. S., Greif, D. M., & Martin, K. A. (2019). Promoters to Study Vascular Smooth Muscle. Arterioscler Thromb Vasc Biol, 39(4), 603-612. doi:10.1161/atvbaha.119.312449
2 .El Agha, E., Moiseenko, A., Kheirollahi, V., De Langhe, S., Crnkovic, S., Kwapiszewska, G., . . . Bellusci, S. (2017). Two-Way Conversion between Lipogenic and Myogenic Fibroblastic Phenotypes Marks the Progression and Resolution of Lung Fibrosis. Cell Stem Cell, 20(2), 261-273.e263. doi:10.1016/j.stem.2016.10.004
3. Chen, Y. C., Gonzalez, M. E., Burman, B., Zhao, X., Anwar, T., Tran, M., . . . Kleer, C. G. (2019). Mesenchymal Stem/Stromal Cell Engulfment Reveals Metastatic Advantage in Breast Cancer. Cell Rep, 27(13), 3916-3926.e3915. doi:10.1016/j.celrep.2019.05.084
Long Name
Actin, Alpha 2, Smooth Muscle, Aorta
Alternate Names
AAT6, ACTA2, Actin alpha 2, ACTSA, ACTVS, alphaSmooth Muscle Actin, MYMY5, SMA
Gene Symbol
ACTA2
Additional alpha-Smooth Muscle Actin Products
Product Documents for alpha-Smooth Muscle Actin Overexpression Lysate
Product Specific Notices for alpha-Smooth Muscle Actin Overexpression Lysate
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.
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