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Follicle-stimulating Hormone(FSH) Elisa Kit for Dianostic Use
Immunoassay for the in vitro quantitative determination of follicle-stimulating hormone in human serum.
FSH (follicle stimulating hormone), together with LH (luteinizing hormone), belongs to the gonadotropin family. FSH and LH regulate and stimulate the growth and function of the gonads (ovaries and testes) synergistically.Like LH, TSH and hCG, FSH is a glycoprotein consisting of two subunits (α- and β‐chains). Its molecular weight is approx. 32000 daltons.In women, the gonadotropins act within the hypothalamus‐pituitary‐ovary regulating circuit to control the menstrual cycle. (1, 2, 3)
FSH and LH are released in pulses from the gonadotropic cells of the anterior pituitary. The levels of the circulating hormones are controlled by steroid hormones via negative feedback to the hypothalamus. In the ovaries FSH, together with LH, stimulates the growth and maturation of the follicle and hence also the biosynthesis of estrogens in the follicles.The FSH level shows a peak at mid-cycle, although this is less marked than with LH. Due to changes in ovarian function and reduced estrogen secretion, high FSH concentrations occur during menopause. (3) In men, FSH serves to induce spermatogoniumdevelopment.Determination of the FSH concentration is used in the elucidation of dysfunctions within the hypothalamus‐pituitary‐gonads system.The determination of FSH in conjunction with LH is utilized for the following indications: congenital diseases with chromosome aberrations, polycystic ovaries (PCO), amenorrhea (causes), and menopausal syndrome. Depressed gonadotropin levels in men occur in azoospermia. (1, 2, 3, 4, 5)
Sandwich principle. Total duration of assay: 80 minutes.
• Sample, Anti-FSH coated microwells and enzyme labeled Anti-FSH are combined.
• During the incubation, FSH presents in the sample is allowed to react simultaneously with the two antibodies, resulting in the FSH molecules being sandwiched between the solid phase and enzyme-linked antibodies.
• After washing, a complex is generated between the solid phase, the FSH within the sample and enzyme-linked antibodies by immunological reactions.
• Substrate solutionis then added and catalyzed by this complex, resulting in a chromogenic reaction. The resulting chromogenic reaction is measured as absorbance.
• The absorbance is proportional to the amount of FSH in the sample.
• Coated Microplate, 8 x 12 strips, 96 wells, pre-coated with mouse monoclonal Anti-FSH.
• Calibrators, 6 vials, 1 ml each, ready to use; Concentrations: 0(A), 5(B), 20(C), 50(D), 100(E) and 200(F) mIU/mL.
• Enzyme Conjugate, 1 vial, 11 mL of HRP(horseradish peroxidase) labeled mouse monoclonal Anti-FSH in Tris-NaCl buffer containing BSA (bovine serum albumin). Contains 0.1% ProClin300® preservative.
• Substrate, 1 vial, 11ml, ready to use, (tetramethylbenzidine) TMB.
• Stop Solution, 1 vial, 6.0 ml of 1 mol/l sulfuric acid.
• Wash Solution Concentrate, 1 vial, 25 ml (40X concentrated), PBS-Tween wash solution.
• IFU, 1 copy.
• Plate Lid: 1 piece.
Materials required (but not provided)
• Microplate reader with 450nm and 620nm wavelength absorbent capability.
• Microplate washer.
• Plate shaker.
• Micropipettes and multichannel micropipettes delivering 50μl with a precision of better than 1.5%.
• Absorbent paper.
• Distilled water
Precautions and warnings
• For in vitro diagnostic use only. For professional use only.
• All products that contain human serum or plasma have been found to be non-reactive for HBsAg, HCV and HIVI/II. But all products should be reared as potential biohazards in use and for disposal.
• Mix the sample in the wells thoroughly by shaking and eliminate the bubbles.
• Conduct the assay away from bad ambient conditions. e.g. ambient air containing high concentration corrosive gas such as sodium hypochlorite acid, alkaline, acetaldehyde and so on, or containing dust.
• Wash the wells completely. Each well must be fully injected with wash solution. The strength of injection, however, is not supposed to be too intense to avoid overflow. In each wash cycle, dry the liquids in each well. Strike the microplate onto absorbent paper to remove residual water droplets. It is recommended to wash the microplate with an automated microplate strip washer.
• Failure to remove adhering solution adequately in the aspiration or decantation wash step(s) may result in poor replication and spurious results.
• Do not use reagents beyond the labeled expiry date.
• Do not mix or use components from kits with different batch codes.
• If more than one plate is used, it is recommended to repeat the calibration curve.
• It is important that the time of reaction in each well is held constant to achieve reproducible results.
• Ensure that the bottom of the plate is clean and dry.
• Ensure that no bubbles are present on the surface of the liquid before reading the plate.
• Thesubstrate and stop solution should be added in the same sequence to eliminate any time deviation during reaction.
• Store at 2-8℃.
• Place unused wells in the zip-lock aluminum foiled pouch and return to 2-8 °C, under which conditions the wells will remain stable for 2 months, or until the labeled expiry date, whichever is earlier.
• Seal and return unused calibrators to 2-8 °C, under which conditions the stability will be retained for 1 month, for longer use, store opened calibrators in aliquots and freeze at -20 °C. Avoid multiple freeze-thaw cycles.
• Seal and return all the other unused reagents to 2-8 °C, under which conditions the stability will be retained for 2 months, or until the labeled expiry date, whichever is earlier.
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