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小鼠/大鼠尿白蛋白定量（Albumin Blue 580 染料结合微量板法）

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Quantifies mouse or rat urinary albumin on a 96-well plate with the Albumin Blue 580 dye-binding reagent and fluorescence readout, following the pinned protocol series-dilution, plating, reading and curve-fitting steps.

Scope and method

Sample：mouse or rat urine

albumin concentration assay

Reagents and materials

Procedure

  1. Prepare working reagent.

    Prepare working reagent.

  2. Prepare standards by serially diluting 200 mg/l standard 1:1 to make 100, 50, 25, 12.5, 6.25 standards.

    Prepare standards by serially diluting 200 mg/l standard 1:1 to make 100, 50, 25, 12.5, 6.25 standards.

  3. Add 25 μl of standard and sample to each well.

    Add 25 μl of standard and sample to each well.

  4. Add 125 μl of working reagent. Read in fluorimeter using 590 nm excitation and 616 nm emission.

    Add 125 μl of working reagent. Read in fluorimeter using 590 nm excitation and 616 nm emission. _**IMPORTANT: **__Make sure not to add any bubbles to the wells when dispensing reagents, this will interfere with reading in the platereader._

  5. Use a polynomial 2nd order curve fit to construct a standard curve. Interpolate the values of the unknowns using the sta

    Use a polynomial 2nd order curve fit to construct a standard curve. Interpolate the values of the unknowns using the standard curve.

Quality control and readouts

标准曲线用二阶多项式拟合；加样避免气泡干扰荧光读数；结果需以同批标准曲线插值。

输出为每样品尿白蛋白浓度（mg/l），按标准曲线插值报告；该读数本身不是白蛋白/肌酐比值，换算比值需另测尿肌酐。

References

  1. UC Davis (AMDCC) Urinary Albumin Excretion protocol, protocols.io v1
  2. Sigma-Aldrich A6414: Albumin from rat serum; biological source rat, UniProt P02770（核对日期2026-10-20）

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