Activin RIIA Overexpression Lysate Summary
Description |
Activin RIIA Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC210227
Accession#: NM_001616
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. |
Gene |
ACVR2A |
Applications/Dilutions
Dilutions |
|
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. |
Theoretical MW |
56 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. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
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.
Alternate Names for Activin RIIA Overexpression Lysate
Background
Activin, a disulfide-linked homodimeric protein is secreted by Sertoli cells in the testis and granulosa cells in the ovary. In early studies, this peptide was thought to be an inhibin and not recognized as a unique compound. Activins and inhibins are members of the TGF-beta superfamily due to amino acid homology with respect to the conservation of 7 of the 9 cysteine residues common to all TGF-beta forms. Activins are homodimers or heterodimers of the various beta subunit isoforms, while inhibins are heterodimers of a unique alpha subunit and one of the various beta subunits. Five beta subunits have been cloned (mammalian betaA, betaB, betaC, betaE, and Xenopus betaD). The activin/inhibin nomenclature reflects the subunit composition of the proteins: activin A (betaA-betaA), activin B (betaB-betaB), activin AB (betaB-betaA), inhibin A (alpha-betaA), and inhibin B (alpha- betaB). Activins have a wide range of biological activities including mesoderm induction, neural cell differentiation, bone remodeling, hematopoiesis, and reproductive physiology. Activins are also involved in growth and differentiation of several tissues from different species. This protein also plays a key role in the production and regulation of hormones such as FSH, LH, GnRH, and ACTH. Activin influences erythropoiesis and the potentiation of erythroid colony formation, oxytocin secretion, paracrine, and autocrine regulation. Similar to other TGF-beta family members, activins exert their biological activities through the effects ot the heterodimeric complex composed of two membrane spanning serine-threonine kinases designated type I and type receptors. Activin type I and type II receptors are distinguished by the level of sequence homology of their kinase domains and other structural and functional features. To date, seven type I and five type II activin receptors have been cloned from mammals, including activin receptor IA, activin receptor IIA, activin receptor IB, and activin receptor IIB. In addition, two splicevariants of activin receptor IIA and five splice variants of activin receptor IIB have been reported. Type I activin receptors are highly conserved and do not bind directly to activin but will associate with the type II receptor-activin complex and initiate signal transduction. Type I activin receptors will also bind with the BMP-2/7-bound BMPR-II and form signaling complexes. Human, mouse, and bovine type IB activin receptors share greater than 98 % homology.
Limitations
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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