Description
Blood potassium plays a crucial role in maintaining normal osmotic pressure, acid-base balance, sugar and protein metabolism, and ensuring normal neuromuscular function. Its concentration is closely related to various important physiological functions. Abnormally high or low levels can disrupt normal physiological activities. Hyperkalemia increases neuromuscular excitability but decreases myocardial excitability, leading to bradycardia. Hypokalemia can cause muscle weakness or even flaccid paralysis, increase myocardial excitability, and result in tachycardia, arrhythmias, or even cardiac arrest during systole.Detection Principle: Potassium ions (K⁺) in serum react with sodium tetraphenylboron to form water-insoluble potassium tetraphenylboron. The turbidity produced is directly proportional to the potassium ion concentration within a certain range. The serum potassium ion content is determined by measuring this turbidity.Applicable Sample: SerumG1501769Component96 TStorageG1501769AExtraction Buffer50 mL2-8℃G1501769BReagentⅠ2.4 mL2-8℃G1501769CReagentⅡ1EA2-8℃. Store in the dark.G1501769DReagentⅢ20 mL2-8℃G1501769EStandard1 mL2-8℃Note: Before formal testing, it is recommended to perform a preliminary test with 2-3 samples expected to have significant differences.User-Prepared Instruments and ReagentsMicroplate reader or visible spectrophotometer (capable of measuring absorbance at 520 nm)96-well plate or micro glass cuvettes, adjustable micropipettes and tipsCentrifugeDeionized water, Concentrated Sulfuric AcidExperimental Procedure1. Reagent PreparationReagent NameReagent PreparationNotesExtraction Buffer/Slightly irritating. Use appropriate personal protective equipment during handling.Working Extraction BufferPrepare before use: Mix Extraction Buffer (mL) and Concentrated Sulfuric Acid (µL) in a 5:12 ratio as needed. Prepare freshly for each use.If precipitate forms, prepare a new batch.Reagent Ⅱ Working SolutionPrepare before use: Add the entire contents of Reagent Ⅰ into the Reagent Ⅱ vial. Mix well. This is the Reagent Ⅱ Working Solution.Can be stored at 4°C protected from light for one week.ReagentⅢPreheat in a 25°C water bath for at least 30 minutes before use./StandardReady-to-use; Equilibrate to room temperature before use.Store at 4°C.2. Sample PreparationSerum Pretreatment: Take a microcentrifuge tube. Sequentially add 50 µL of serum and 450 µL of Working Extraction Buffer. Mix thoroughly. Centrifuge at 8,000 rpm for 10 minutes at room temperature (approximately 25°C). Collect the supernatant for assay.3. Assay Steps3.1 Preheat the microplate reader or visible spectrophotometer for at least 30 minutes. Set the wavelength to 520 nm. For spectrophotometers, zero the instrument with deionized water.3.2 Assay Procedure (perform in a 96-well plate or micro glass cuvette):ReagentBlank Well (µL)Standard Well (µL)Test Well (µL)Deionized Water4000Standard0400Supernatant0040Reagent Ⅱ Working Solution202020Mix well and let stand for 5 minutes.ReagentⅢ1401401403.3 Mix well after addition. Measure the absorbance at 520 nm. Record the absorbance of the Blank well as A blank, the Standard well as A standard, and the Test well as A test. Note:The Blank and Standard tubes only need to be set up 1-2 times.It is recommended to perform a preliminary test with 2-3 samples expected to have significant differences before the formal experiment. If A <sub> test </sub> is less than 0.02, consider increasing the sample volume appropriately. If A <sub> test </sub> is greater than 1.1, further dilute the sample with Working Extraction Buffer (multiply the result by the dilution factor) or reduce the sample volume used for extraction.4. Calculation of ResultsNote: We provide both the derived formula and a simplified formula. They are equivalent. It is recommended to use the simplified formula in bold for final calculation.Blood Potassium Concentration (mmol/L) = [C Standard × (A test - A blank ) ÷ (A standard - A blank )] × n = 5 × (A test - A blank ) ÷ (A standard - A blank )Parameter Definitions:C Standard : 0.5 mmol/Ln: Sample dilution factor (10)Precautions1. Before formal testing, it is recommended to perform a preliminary test with 2-3 samples expected to have significant differences.2. Biochemical reagents are generally irritating and potentially biologically toxic. For your safety and health, please use appropriate biosafety precautions throughout the experiment, including wearing lab coats, masks, gloves, and head covers. Perform experiments in a fume hood or biosafety cabinet.3. This product is for research use only. Not for use in clinical diagnosis