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| DNA Sequencing | The corresponding genes were synthesized including a se- quence encoding a C-terminal His6 tag and cloned into a pIRES2-AcGFP1 plasmid (Clontech, Mountain View, CA, USA) using the restriction enzymes XhOI and BamHI; plas- mids were verified by sequencing (made by GenScript, Piscataway, NJ, USA) CHO-K1 cells (ATCC, Teddington, UK) cultured in F-12 K Kaighn's medium with 10% FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml) (Gibco/Invitrogen, Grand Island, NY, USA) were transfected with LAP or mock plasmids using lipofectamine 2000 (Gibco).... Wells coated with anti-His6 mAb (GenScript) were incubated with LAP1, -2 and − 3 CHO cell samples, followed by detection with biotinylated anti-LAP1 mAbs and SA-HRP. | Get A Quote |
Human Transforming Growth Factor (TGF)-β1， one of three TGF-β isoforms， is a pleotropic cytokine critical for many physiological and immunological processes. TGF-β1 is secreted in a latent form， linked to Latency Associated Protein (LAP). Analysis of Latent TGF-β1 by TGF-β1 ELISA requires dissociation of TGF-β1 from LAP， e.g. by acidification of samples. The ELISA then measures total TGF-β1， equivalent to dissociated Latent TGF-β1 plus any free TGF-β1 present prior to acidification. Evolutionary conservation of TGF-β1 across mammals also renders TGF-β1 ELISAs reactive with TGF-β1 in bovine serum often used in human cell cultures. To enable a direct analysis of Latent TGF-β1， monoclonal ... More
