Ultra-sensitive HumanTSH ELISA Kit

serum

All reagentsprovided are stored at 2-8oC. Refer to expiration date on the label. 96 tests 1. MICROTITER PLATE:96 wells Pre-coated withanti-human TSH monoclonal antibody. 2. CONJUGATE: 12 mL Anti-human TSHpolyclonal antibody conjugated to horseradish peroxidase (HRP) with preservative.Ready-to-use. 3. STANDARD - 12uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 12 uIU/mL afterreconstitution. 4. STANDARD - 6uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 6 uIU/mL afterreconstitution. 5. STANDARD- 2 uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 2 uIU/mLafter reconstitution. 6. STANDARD – 0.8uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 0.8 uIU/mL afterreconstitution. 7. STANDARD – 0.2uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 0.2 uIU/mLafter reconstitution. 8. STANDARD- 0 uIU/mL: 1 vial Lyophilized bufferedprotein base with preservative that will contain 0 uIU/mL of human TSH after reconstitution. 9. UBSTRATE A: 10mL Buffered solutionwith H2O2. 10. UBSTRATE B: 10mL Buffered solutionwith TMB. 11. TOP SOLUTION: 7mL 2N Sulphuric Acid (H2SO4).Caution: Caustic Material!

1. PREPARATION OFREAGENTS Remove all kitreagents from refrigerator and allow them to reach room temperature (20-25oC).Prepare the following reagents as indicated below. Mix thoroughly by gently swirlingbefore pipetting. Avoid foaming. 1) TSH Standards:Reconstitute each TSH Standard vial with 0.6 mL of deionized or distilledwater. Allow each solution to sit for at least 15 minutes with gentleagitation. The TSH standard stock solutions are stable at 4C for 3 months.Avoid freezethaw cycles. 2) SubstrateSolution: Substrate A and Substrate B should be mixed together in equal volumesup to 15 minutes before use. Refer to the table below for correct amounts of SubstrateSolution to prepare. 2. ASSAY PROCEDURE 1) Prepare all TSHStandards before starting assay procedure. 2) First, secure thedesired number of coated wells in the holder, then add 100 uL of Standards orsamples to the appropriate well of the antibody pre-coated Microtiter Plate. 3) Add 100uL of Conjugateinto each well. COMPLETE MIXING IN THIS STEP IS IMPORTANT. Cover and incubatefor 3 hours at 37oC. 4) 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 withde-ionized or distilled water, and then aspirate contents of the plate into asink or proper waste container. Repeat this procedure four more times for atotal of FIVE washes. After final wash, invert plate, and blot dry by hittingplate onto absorbent paper or paper towels until no moisture appears. Note:Hold the sides of the plate frame firmly when washing the plate to assurethat all strips remain securely in frame. b. AutomatedWashing: Aspirate all wells, then wash plates FIVE times using distilled orde-ionized water. Always adjust your washer to aspirate 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. 5) Add 100uL ofSubstrate into each well. Cover and incubate for 15 minutes at 37oC. 6) Add 50uL of StopSolution to each well. Mix well. 7) Read the OpticalDensity (O.D.) at 450 nm using a microtiter plate reader within 30 minutes.

1.SENSITIVITY Theminimal detectable concentration of TSH by this assay is estimated to be 0.05uIU/mL. 2.SPECIFICITY This kitexhibits no detectable cross-reaction with human FSH, LH, Prolactin, or CG.hGH can be detected in this assay. 3.CALIBRATION T Thisimmunoassay is calibrated against WHO, 2nd IRP, 80/558. 4. HOOKEFFECT In thisassay, no hook effect is observed up to 2000 uIU/ml. 5.EXPECTED NORMAL VALUES Eachlaboratory must establish its own normal ranges based on patient population. Theresults provided below are based on random normal adult blood specimens. Normal Hyperthyroid N 16046 Mean TSH(uIU/mL) 1.6 - Range0.4-7.0 0.07-0.14