Vitellogenin (VTG) ELISA Kit

Antigen: Vitellogenin (VTG)

Synonyms: vg4, wu:fb59b02, wu:fb59d07, wu:fb59e10, si:rp71-23d18.3, vg5, MGC114012, wu:fi49a01, zgc:114012, si:rp71-23d18.5, vtg b1, Vg, GB13999, vg6, wu:fb62e09, si:rp71-23d18.1, vg7, MGC158513, wu:fb62f05, si:dkey-4c23.1, vtga2, VTG, vtg1

Reactivity: Carp

Application: ELISA

Catalog no.: ABIN368675

Additional Information

Alternative name: Vitellogenin

Description: Vitellogenin (Vg) is a precurser of egg yolk and detected female serum near ovulation. Vg is not detected in male serum, but treated with 17beta-estradiol, Vg is detectable in male serum. Carp Vg ELISA kit is constructed as a sandwich ELISA format using anti carp Vg antibodies and titrate serum carp Vg.

Sensitivity: Dynamic range: 7.8-500 ng/mL. Recovery of exogenous vitellogenin Plasma: Av. 95.6%

Synonyms: vg4, wu:fb59b02, wu:fb59d07, wu:fb59e10, si:rp71-23d18.3, vg5, MGC114012, wu:fi49a01, zgc:114012, si:rp71-23d18.5, vtg b1, Vg, GB13999, vg6, wu:fb62e09, si:rp71-23d18.1, vg7, MGC158513, wu:fb62f05, si:dkey-4c23.1, vtga2, VTG, vtg1

Application Details

Principle: This kit is constructed using the antibodies specific to carp Vg , as both capture and secondary antibodies of sandwich ELISA format. During the assay, carp Vg in the samples is captured by the antibody, immobilized on the surface of microplate wells, and simultaneously reacts with the horseradish peroxidase (HRP) labeled antibody. Quantitation of carp Vg in samples can be made by measurement of HRP activity of the complex, which is exponential to the quantity of carp Vg.

Protocol: (1) Preparation of reagents to be used: (a) Antibody-coated microtiter plate: The 96 well microplate is coated with antibody to carp Vg and is ready-to-use. Add 300uL of well. Wash solution for 10 to 60 minutes at room temperature. (b) Standard Carp Vg: This standard concentration is 1ug/ml. In order to make the standard dilution 1ug/ml carp Vg solution is diluted in two serial dilutions from 7.8 ng/mL to 500 ng/mL with dilution solution. (c) HRP-conjugated anti Carp Vg antibody: The stock HRP-conjugated anti carp Vg antibody can be prepared for 48 well reaction upon dilution of 0.01mL stock solution with 5 mL Dilution solution just before use. (d) Substrate solution: One OPD tablet is dissolved in 6.5 mL substrate buffer. (e) Wash solution: Make sure that the solution does not contain any crystalized material, upon reaching room temperature. Working solution is prepared by dilution of 30mL of the stock wash solution with distilled deionized water to 570mL. This solution is stable for 14 days when stored in a refrigerator. (2) Assay procedure: (a) Discard the wash solution from the wells and dispence 100uL standard diluent into zero blank wells. (b) In the assay each concentration of standard solution and serum sample dispence 100uL into each well. Incubate at room tempelature for 2 hours. (c) Discard the solution from the wells, and then wash 3 times with 300uL of washsolution. (d) Dispence 100uL HRP labeled anti carp Vg antibody into the wells, and incubate at room temperature for one hour. (e) Discard the solution from the wells, and then wash 5 times with 300uL/well of with wash solution. (f) Dispense 100uL of substrate solution into the wells, and incubate at room temperature for 15 minutes. (g) Dispence 50uL of stop solution and read absorbance with a microplate reader at 490 or 492nm.

Application Notes: 1. Duplicate assay should be recomended for each sample. 2. The standard curve should be made during each assay. 3. Serum is diluted with standerd diluent more than 5 dilutions. 4. HRP-conjugated anti-Carp Vg antibody is not stable under sun light, so that do not leave it for long time under sun light. 5. Dissolved carp standerd Vg should not store at room temperature. 6. The appalatus for making coloring solution should be clean one. 7. OPD (o-phenylendiamin) is harmful, so that take good care to it. 8. The kit is constructed with well-adjusted combination for each lot. Replace combination among different lots may cause unexpected results. 9. Stop solution, 1N H2SO4, is strong acid, so that take good care to it. 10. This kit is only for research use. Do not use for medicine or any other things. 11. When using the reagents, take care to avoid them from touching to skin, mucous membrane, clothes, and getting into eye. 12. If the reagents happen to get into eye or mouth, wash out them and go to see a doctor if you need. 13. After using the kit, wash your hand very carefully. 14. If you find that the packages of the reagents are broken or something trouble, do not use them. 15. When you store the reagents, make sure to avoid them from vaporizing, falling down. 16. After using the reagents, the packages should be discarded under established rule. 17. We can not guarantee for any other usage of the packages and the accompaniments.

Components: (1) Antibody-coated microtiter plate (8wellsx12) 1 plate. (2) The standard, Carp Vg 1ug/ml 300uLx2vials. (3) Dilution solution 30 mL(4) HRP-conjugated anti Carp Vg antibody (x500) 30 uL. (5) Substrate buffer 15 mL. (6) OPD (o-phenylendiamin) tablets 2 tab. (7) Wash solution (x20) 30 mL (8) Stop solution 6 mL

Material not included: (1) A microplate reader. (2) A micropipet. (3) A microplate washer

Storage: Store at -80 degrees Celsius

Research Area: Hormones

Restrictions: For Research Use only

Images

The standard curve of carp Vg is obtained by measurement of the serial dilution of standard carp Vg solution with the kit

Reproducibility of results