VTG | Sole vitellogenin
AS06 127 | clonality: polyclonal | host: rabbit | reactivity: sole
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|Recommended dilution||1 : 5 000 on sole serum (ELISA), 1 : 5 000 (WB)|
|Expected | apparent MW||
ca. 200 kDa
|Confirmed reactivity||Senegalese sole, Dicentrarchus labrax (sea bass), Sparus aurata (seabream)|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
The developed VTG ELISA, using these VTG and AbVTG, has been validated for Senegalese sole, sea bass (Dicentrarchus labrax) and seabream (Sparus aurata), which all gave parallel displacement curves in the assay. Probably, plasmas from several other fish species displace parallel and can also be used in the assay, although it has to be validated for each case.
|Selected references||Guzman et al. (2008) Vitellogenin, steroid plasma levels and spawning performance of cultured female Senegalese sole (Solea senegalensis).Gen and Comp Endocrinology 156: 285-297.|
STD: Protein standard (molecular weights of the proteins are indicated on the left) (1) plasma from male sole (load 0.11 ul of plasma),(2) plasma from vitellogenic female sole (load 0.11 ul of plasma), (3) plasma from estradiol-treated male sole (load 0.08 ul of plasma,(4) VTG precipitate (load 0.83 ul of precipitate),(5) purified VTG (load 2 ul of purified preparation, corresponding to around 0.14 ug and spawning performance in cultured VTG) were separated on SDS-PAGE 7.5% resolving gel, 4% stacking gel. Samples were denatured in SDS and b- mercaptoethanol and treated 4 min 95 oC before loading. Following gel electrophoresis proteinswere transfered to PVDF Membrane (Inmobilon-P, Millipore) for 2 h. Blots were blocked in TBST containing 2% non-fat dry milk. Blots were incubated in primary antibody at a dilution 1: 40 000, followed by incubation with secondary antibodies (GAR-HRP, BioRad, dilution 1:2,000) and chemiluminiscence development (Luminol Reagent, Santa Cruz Biotechnology).
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