Human FSH ELISA Kit

RUO, This Human FSH ELISAKit is to be used for the in vitro quantitative determination of humanfollicle stimulating hormone (FSH) concentrations in serum. This kit isintended FOR LABORATORY RESEARCH USE ONLY and is not for use in diagnostic ortherapeutic procedures.

Follicle Stimulating Hormone

1. Single ormulti-channel precision pipettes with disposable tips: 10-100uL and50-200uL forrunning the assay. 2. Pipettes: 1 mL, 5mL 10 mL, and 25 mL for reagent preparation. 3. Multi-channelpipette reservoir or equivalent reagent container. 4. Test tubes andracks. 5. Polypropylenetubes or containers (25 mL). 6. Incubator (37oC). 7. Microtiter platereader (450nm +/- 2nm) 8. Automaticmicrotiter plate washer or squirt bottle. 9. Sodiumhypochlorite solution, 5.25% (household liquid bleach). 10. Deionized ordistilled water. 11. Plastic platecover. 12. Disposablegloves. 13. Absorbent paper.

1.Reagent And Sample Preparation Remove all kitreagents from refrigerator and allow them to reach room temperature(20-25oC). Prepare the following reagents as indicated below. Mix thoroughlyby gently swirling before pipetting. Avoid foaming. 1) FSH Standards:Reconstitute each FSH Standard vial with 0.6 mL of distilled or deionizedwater. Allow each solution to sit for at least 15 minutes with gentleagitation. The FSH standard stock solutions are stable at 4C for 3 months.Avoid freeze-thaw cycles. 2) SubstrateSolution: Substrate A and Substrate B should be mixed together in equalvolumes up to 15 minutes before use. Refer to the table below for correctamounts of Substrate Solution to prepare. 2. Assay Steps 1) Prepare all FSHStandards before starting assay procedure. 2) First secure thedesired number of coated wells in the holder, then add 50 uL ofStandards or samples to the appropriate well of the antibody pre-coatedMicrotiter Plate. 3) Add 100uL ofConjugate into each well. COMPLETE MIXING IN THIS STEP IS IMPORTANT. Coverand incubate for 1 hour at 37oC. 4) Prepare SubstrateSolution no more than 15 minutes before end of incubation (see Preparation ofReagents). 5) Wash theMicrotiter Plate using one of the specified methods indicated below: a. Manual Washing:Remove incubation mixture by aspirating contents of the plate into a sink orproper waste container. Using a squirt bottle, fill each well completely withdistilled or de-ionized water then aspirate contents of the plate into a sink orproper waste container. Repeat this procedure four more times for a total ofFIVE washes. After final wash, invert plate, and blot dry by hitting plateonto absorbent paper or paper towels until no moisture appears. Note: Holdthe sides of the plate frame firmly when washing the plate to assure that allstrips remain securely in frame. b. AutomatedWashing: Aspirate all wells, then wash plates FIVE times using distilled orde-ionized water. Always adjust your washer to apirate as much liquid aspossible and set fill volume at 350 uL/well/wash (range:350-400 uL). Afterfinal wash, invert plate, and blot dry by hitting plate onto absorbent paperor paper towels until no moisture appears. It is recommended that the washerbe set for soaking time of 10 seconds or shaking time of 5 seconds betweenwashes. 6) Add 100 uLSubstrate Solution to each well. Cover and incubate for 15 minutes at 37oC. 7) Add 50uL of StopSolution to each well. Mix well. 8) Read the OpticalDensity (O.D.) at 450 nm using a microtiter plate reader within 30 minutes.

1. SENSITIVITY The minimaldetectable concentration of FSH by this assay is estimated to be 1.5 mIU/mL. 2. SPECIFICITY This kit exhibits nodetectable cross-reaction with LH, HCG, TSH, and Prolactin. hGH can bedetected in this assay. 3. CALIBRATION This immunoassay iscalibrated against NIBSC/WHO, 3rd IS, HMG. 4. HOOK EFFECT In this assay, nohook effect is observed up to 5000 mIU/mL. 5. EXPECTED NORMALVALUES Each laboratory mustestablish its own normal range based on patient population. The resultsprovided below are based on random selected out-patient clinical laboratorysamples