Vitellogenin (VTG) ELISA Kit

Antigen: Vitellogenin (VTG)

Synonyme: 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

Reaktivität: Japanische Wachtel

Applikation: ELISA

Produktnummer: ABIN368676

Produktbeschreibung

Weitere Bezeichnung: Vitellogenin

Beschreibung: In recent years, influence of Endocrine Disrupting Chemicals (EDC) upon living organisms has been one of the hot topics in the environmental science. However, evaluation methods of chemical compound species that may have EDC effects has not yet been established, and it is absolutely necessary to take action to complete such a task. Vitellogenin (Vg), a precursor of yolk protein, is a female-specific protein that is present only in the blood of higher oviparous vertebrate organisms. Vitellogenin is normally expressed in the liver under regulation of estrogen (female sex hormone) during an egg formation process, and when it reaches a growing oarian follicle via blood stream it contributes to the egg yolk formation. Since estrogen is known to have an effect upon male species to induce unusual expression of Vg, an ELISA screening method detecting Vg in EDC exposed male animals is the most reliable bio-screening assay system.

Spezifität: Recovery test: In the recovery study, recoveries 99.8% and 98.3% were obtained for 8 times dilutions of the serum sample.

Synonyme: 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

Anwendungen

Prinzip: This kit is based on the ELISA method which uses two different specific anti-Japanese Quail Vg antibodies. Anti-Japanese Quail monoclonal antibody is coated on the microplate, so that Vg in the sample and standard solution is captured. Reacting with anti-Japanese Quail polyclonal antibody and HRP-conjugated anti-Rabbit IgG, the complex of coated-antibody antigen-antibody is formed. Finally, Vg concentrations are estimated according to the coloring intensity developed with enzymatic reaction.

Protokoll: (1) Preparation of reagents to be used: (a) Wash solution. Make sure that the solution does not contain any crystallized material at room temperature. Working solution is prepared by dilution of 30 mL of the stock wash solution with distilled deionized water to 570 mL. This solution is stable for 14 days when stored in a refrigerator. (b) Antibody-coated microtiter plate: Add wash solution 300uL to each well and wait another 10 to 60 minutes. (c) Standard dilution: Dilute the concentrated standard solution (200 ng/mL ) to make 100 50 25 12.5 6.25 3.13 1.56 0.78 ng/mL with dilution solution. (d) Anti-Japanese Quail Lipovitellin polyclonal antibody (x100). Dilute the stock solution of 50uL anti-Japanese Quail Lipovitellin polyclonal antibody with 5 mL of Dilution solution for 96 well reaction. Make solution just before use. (e) HRP-conjugated anti-Rabbit IgG (x100) Dilute the stock solution of 50uL HRP-conjugated anti-Rabbit IgG with 5 mL Dilution solution for 96 well reaction just before use. (f) Coloring solution: Dissolve two OPD tablets with 13 mL of Substrate buffer. (2) Preparation of samples to be used: The serum of Japanese Quail should be diluted more than 8 times with Dilution solution. (3) Assay procedure: (a) Discard the wash solution from the wells completely, but do not keep wells in dry condition for reliable data. (b) Apply 50uL of each concentration of standard solution and serum samples into the wells and incubate for 1 hour. (c) After the incubation, discard the reaction solution and wash with 300uL wash solution. Repeat this washing procedure another 2 times. (d) Apply 50uL anti-Japanese Quail Lipovitellin polyclonal antibody and incubate for 1 hour. (e) After the incubation, wash the wells 3 times with wash solution (See (c)). (f) Apply 50uL HRP-conjugated anti-Rabbit IgG and incubate for 1 hour. Substrate buffer should be restored to room temperature from refrigerator at this time. (g) After the incubation, wash the wells 3 times with wash solution (See (c)). (h) Apply 100 Coloring solution into each well for 5 minutes in room temperature. (i) Apply 50uL of Stop solution for stopping the enzymatic reaction. (j) Read absorbance with a microplate reader at 490nm or 492nm. (k) Estimate the Japanese Quail Vitellogenin concentration of each sample using the standard curve.

Applikationshinweise: (a) The Reagents should be stored at recommended temperature. (b) Do not use the reagents which is expired the date of usage. (c) The serum of Japanese Quail should be diluted more than 8 times with Dilution solution. (d) Do not leave the standard, Japanese Quail Vg, for long time under room temperature. (e) Since HRP-conjugated anti-Rabbit IgG is not stable under sun light, do not leave it for long time under sun light. (f) The glassware for making coloring solution should be clean. (g) Since OPD (o-phenylendiamine) is harmful, handle with care. (h) Since Stop solution,1N H2SO4, is strong acid, handle with care. (i) The kit is constructed with well-adjusted combination in each lot. Replaced combination among different lots may cause unexpected results. (j) This kit is only for research use. Do not use for medicinal or any other purposees. (k) When using the reagents, take care to avoid them from touching to skin, mucous membrane, clothes, and getting into eye. (k) If the reagents happen to get into eye or mouth, wash out them and consult a doctor if you need. (k) After using the kit, wash your hand very carefully. (l) If you find that the packages of the reagents are broken or something wrong, do not use them. (m) When you store the reagents, make sure to avoid them from vaporizing, falling down. (n) After using the reagents, the packages should be discarded under the established rule. (o) We do not guarantee the quality of the packages and accompaniments if not used according this direction.

Bestandteile: (1) Antibody-coated microtiter plate (8 wellsx12) 1 plate. (2) The standard, Japanese Quail Vg 200ng/mL 300uL x 2. (3) Dilution solution 30mL x 2. (4) Anti-Japanese Quail Lipovitellin polyclonal antibody (x100) 80uL x 1. (5) HRP-conjugated anti-Rabbit IgG (x100) 80uL x 1. (6) OPD (o-phenylendiamine) tablets 2 tab. (7) Substrate buffer 15 mL x 1. (8) Stop solution 6 mL x 1. (9) Wash solution (x20) 30 mL x 1

Benötigtes Material: (1) A microplate reader. (2) A micropipet. (3) A microplate washer

Lagerung: Store at -80 degrees Celsius

Forschungsgebiet: Hormone

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

Standard Curve

Reproducibility of results