| Usage | I. Supplies to be Provided: 1. Microplate reader (capable of measuring absorbance at 450 nm) 2. Multichannel pipette or automated plate washer 3. Incubator, refrigerated centrifuge 4. Adjustable pipette and tips 5. Deionized water II. Reagent Preparation: Standard Diluent: Ready-to-use; equilibrate to room temperature before use; store at 2-8°C. Standard Diluent is used to dilute standards. 1× Assay Buffer: Before use, equilibrate to room temperature and dilute Assay Buffer (10×) with deionized water at a ratio of 1:10 to obtain 1× Assay Buffer. Mix gently to avoid foaming. Store at 2-8°C. This solution is stable for 30 days. 1× Assay Buffer is used to dilute serum and plasma samples. HRP-conjugated Human TREM2 Detect Antibody (100×).
Human TREM2 Standard: Reconstitute the Human TREM2 Standard with deionized water in the volume indicated on the standard label to yield a 20 ng/mL standard.
Before dilution, gently agitate the standard for at least 15 minutes. After reconstitution, the standard can be stored at -20°C for up to 1 month. Reconstitute once and discard.
1× HRP-conjugated Human TREM2 Detect Antibody: Mix thoroughly before dilution. Based on the required volume of standard and sample, dilute HRP-conjugated Human TREM2 Detect Antibody (100×) 1:100 in a clean plastic tube with 1× Assay Buffer. 1× HRP-conjugated Human TREM2 Detect Antibody should be used within 30 minutes of dilution. HRP Substrate (TMB): Ready-to-use; equilibrate to room temperature before use; store at 2-8°C in the dark. Stop Solution: Ready-to-use; equilibrate to room temperature before use; store at 2-8°C. 1× Wash Buffer: Equilibrate to room temperature before use. Dilute 20× Wash Buffer 1:20 with deionized water to create 1× Wash Buffer. Mix gently to avoid foaming and store at room temperature. This solution is stable for 30 days.
Standard Curve Setup: Dilute the 20 ng/mL standard with Standard Diluent to 10, 5, 2.5, 1.25, 0.63, 0.31, 0.16, and 0 ng/mL Human TREM2 standards as shown in the table below.
| Serial number | Standard Diluent | Standard Diluent
Volume (μL) | Standard concentration (ng/mL) | | Std.1 | 230 μL of 20 ng/mL | 230 | 10 | | Std.2 | 230 μL of Std.1 10 ng/mL) | 230 | 5 | | Std.3 | 230 μL of Std.2 (5 ng/mL) | 230 | 2.5 | | Std.4 | 230 μL of Std.3 (2.5 ng/mL) | 230 | 1.25 | | Std.5 | 230 μL of Std.4 (1.25 ng/mL) | 230 | 0.63 | | Std.6 | 230 μL of Std.5 (0.63 ng/mL) | 230 | 0.31 | | Std.7 | 230 μL of Std.6 (0.31 ng/mL) | | | Kit components | 48T | 96T | Save | | Human TREM2 Microplate | 48 wells | 96 wells | 2-8℃ | | Human TREM2 Standard (lyophilized) | 1 | 2 | 2-8℃ | | Standard Diluent | 5mL | 5mL | 2-8℃ | | Assay Buffer (10×) | 5mL | 5mL | 2-8℃ | | 1 | 1 | 2-8℃ | | HRP Substrate (TMB) | 6mL | 11mL | 2-8℃, away from light | | Stop Solution | 11mL | 11mL | 2-8℃ | | Wash Buffer (20×) | 50mL | 50mL | 2-8℃ | | Plate Covers | 6 | 6 | Room temperature |
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