|Detection of Mouse Lefty-2 by Western Blot. Western blot shows lysates of P19 mouse embryonal carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Mouse Lefty-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7648) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Lefty-2 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Lefty-2 in D3 Mouse Embryonic Stem Cells. Lefty-2 was detected in immersion fixed D3 mouse embryonic stem cells untreated (lower panel) or stimulated (upper panel) with recombinant human/mouse/rat Activin A (Catalog # 338-AC) using Sheep Anti-Mouse Lefty-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7648) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm of Activin A-stimulated cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
Lefty-2 (Left-right determination factor 2; also Lefty-B [in human]) is a atypical member of the TGF-beta family of proteins. It is expressed during early embryogenesis in both the primitive streak and left-side lateral plate mesoderm. In the adult, Lefty-2 appears in oviduct epithelium. Lefty-2 acts in a manner reminiscent of that for Chordin and Noggin, and it is assumed that Lefty-2 is an antagonist of BMP activity. Notably, Lefty-2 and Nodal are likely under the control of Lefty-1, and thus all three molecules would appear to contribute to the creation of a left side-type body plan. Mouse Lefty-2 is synthesized as a 368 amino acid (aa) preproprecursor. It contains a 21 aa signal sequence, plus a 347 aa, 41-42 kDa bioactive proprecursor that may undergo proteolytic processing at one of two downstream cleavage sites. If cleavage occurs after Arg77, the resulting 33-34 kDa mature form (aa 78-368) is biologically inactive; if cleavage occurs after Arg135, the resulting 27-28 kDa mature form (aa 136-368) is biologically active. Lefty-2 is not a covalent homodimer and has been suggested to act as a monomer. Over aa 78-368, mouse Lefty-2 shares 94%, 83% and 95% aa sequence identity with rat Lefty-2, human Lefty-B and mouse Lefty-1, respectively.
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