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At,,70%,,ammonium,,sulfate,,saturation,,,almost,,all,,contaminating,,protei ns,,were,,removed,,except,,for,,a,,20-kDa,,protein,,(Figure,,1,,,lanes,,6,, and,,7,,,arrowed,,band).,,Peters,,T,,Jr.,,%10152025303335404550556065707580 859095100.,,Figure,,1,,:,,SDS-PAGE,,analysis,,of,,BSA,,purification,,from,, serum,,by,,ammonium,,sulfate,,precipitation.,,803877117157.,,J,,,Protein,,, Chem.,,,BSA,,,started,,,eluting,,,from,,,the,,,column,,,at,,,a,,,NaCl,,,con centration,,,of,,,150,,,mM,,,but,,,not,,,significantly,,,until,,,250,,,mM,, ,(Figure,,,2).,,,Hepatology.,,Acetylation,,of,,BSA,,to,,inactivate,,DNase,, limits,,its,,use,,as,,a,,protein,,standard,,due,,to,,interference,,with,,co lor,,development,,in,,assays,,such,,as,,Lowry's.,,
It,,has,,been,,demonstrated,,that,,parameters,,such,,as,,sample,,concentrat ion,,,sample,,size,,and,,flow,,rate,,significantly,,influence,,chromatograp hic,,separation,,of,,BSA,,and,,IgG,,[28].,,The,,,,blotted,,,,protein,,,,sam ples,,,,were,,,,probed,,,,with,,,,a,,,,HRP-conjugated,,,,secondary,,,,antib ody,,,,that,,,,reacts,,,,with,,,,the,,,,complete,,,,bovine,,,,IgG,,,,protei n,,,,as,,,,well,,,,as,,,,the,,,,light,,,,chains,,,,of,,,,other,,,,bovine,,, ,immunoglobulins.,,,,In,,,,Putnam,,,,FW,,,,(Ed.),,,,,The,,,,plasma,,,,prote ins:,,,,structure,,,,,function,,,,and,,,,genetic,,,,control,,,,,1975,,,,,1: ,,,,141,,,,,147.,,,,It,,is,,a,,specific,,case,,of,,a,,more,,general,,techni que,,known,,as,,salting,,out.,,Finally,,,,,we,,,,tested,,,,the,,,,purified, ,,,BSA,,,,for,,,,the,,,,presence,,,,of,,,,endotoxin,,,,using,,,,the,,,,Toxi nSensor,,,,Chromogenic,,,,LAL,,,,assay,,,,kit,,,,(GenScript,,,,,Piscataway, ,,,,NJ,,,,,USA),,,,and,,,,found,,,,that,,,,it,,,,contained,,,,4,,,,EU/mL,,, ,endotoxin,,,,(result,,,,not,,,,shown).,,,,As,,,,the,,,,salt,,,,concentrati on,,,,(ionic,,,,strength),,,,is,,,,increased,,,,further,,,,,the,,,,solubili ty,,,,of,,,,the,,,,protein,,,,begins,,,,to,,,,decrease.,,,,Biotechniques.,, ,Heat,,denaturation,,of,,bovine,,serum,,albumin,,in,,alkaline,,pH,,region., ,
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