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VTG | Sole vitellogenin

AS06 127  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: sole

VTG | Sole vitellogenin  in the group Human/Animal Antibodies / Fish Proteins at Agrisera AB (Antibodies for research) (AS06 127)

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product information
Background

Vitellogen (VTG) is a glucolipoprotein yolk precursor produced by all oviparous animals. This protein is female-specific and its expression is under hormonal control (estrogen). However, in the presence of Endocrine Disrupting Chemicals (EDC's), males can express the VgTG gene in a dose dependent manner. The use of VTG gene expression in male fish can be used as a molecular marker of exposure to estrogenic EDC's.

Immunogen

native vitellogenin purified from plasma of estradiol induced male of Senegalese sole (Solea senegalensis)

Host Rabbit
Clonality Polyclonal
Clone
Purity Serum
Format Lyophilized
Quantity 200 ĩl
Reconstitution For reconstitution add 200 ĩl of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications ELISA (ELISA), Western blot (WB)
Related products

collection of antibodies to fish proteins

Secondary antibodies

Additional information

VTG can be purified using following methodology: 
Mañanós et al. (1994). Sea bass (Dicentrarchus labrax L.) vitellogenin. I—Induction, purification and partial characterization. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, Vol 107 (2): 205-216.
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.

application information
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)
Predicted reactivity
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information

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.

application example western blot using anti-VTG antibodies
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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