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 - 40uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 40 uIU/mL afterreconstitution. 4. STANDARD - 20uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 20 uIU/mL afterreconstitution. 5. STANDARD- 10 uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 10 uIU/mLafter reconstitution. 6. STANDARD - 5uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain 5 uIU/mL afterreconstitution. 7. STANDARD - 1uIU/mL: 1 vial Lyophilized humanTSH in a buffered protein base with preservative that will contain1 uIU/mL afterreconstitution. 8. STANDARD- 0 uIU/mL: 1 vial Lyophilized bufferedprotein base with preservative that will contain 0 uIU/mL of human TSH afterreconstitution. 9. UBSTRATE A: 10mL Buffered solutionwith H2O2. 10. UBSTRATE B: 10mL Buffered solution withTMB. 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: Reconstituteeach TSH Standard vial with 0.6 mL of deionized or distilled water. Alloweach solution to sit for at least 15 minutes with gentle agitation. The TSHstandard stock solutions are stable at 4C for 3 months. Avoid freezethaw cycles. 2) Substrate Solution:Substrate A and Substrate B should be mixed together in equal volumes up to15 minutes before use. Refer to the table below for correct amounts ofSubstrate Solution 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 50 uL of Standards orSamples to the appropriate well of the antibody pre-coated Microtiter Plate. 3) Add 2 drops or100 uL of Conjugate to each well. Mix well. Complete mixing in this step isimportant. Cover and incubate for 2 hours 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 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. 6) Add 100uL ofSubstrate into each well. Cover and incubate for 15 minutes at 37oC. 7) Add 100 uL ofStop Solution to each well. Mix well. 8) 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 0.2uIU/mL. 2. SPECIFICITY This kitexhibits no detectable cross-reaction with human FSH, LH, Prolactin, or CG. HumanGH can be detected in this assay. 3.CALIBRATION 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 160 random normal adult blood specimens. Sample(n) Mean TSH (uIU/ml) Range(uIU/ml) 1601.6 0.4– 7.0