PAP Prostatic Acid Phosphatase

PAP Prostatic Acid Phosphatase

1.        Bring all reagents and samples to room temperature (20oC - 25oC) and shake gently before beginning the test. Have all reagents and samples ready before the start of the assay. Once the test is begun it must be performed without any interruption to get the most reliable and consistent results.
2.        Use new disposable tips for each specimen.
1.        Secure the desired number of coated wells in the holder.
2.        Dispense 25 ul of references, controls or serum samples into the appropriate wells.
3.        Dispense 100 ul of enzyme conjugate into wells. Incubate for 30 minutes at room temperature.
4.        Remove incubation mixture and rinse the wells 5 times with running water.
5.        Dispense 100 ul of Solution A and Solution B into each Well and incubate for 10 minutes at room temperature.
6. Stop reaction by adding 50 ul of 1 N H2SO4 to each well and read absorbance at 450 nm with ELISA reader against Blank well (only solution A and solution B)

1.        Plot the concentration (X) of each reference standards against its absorbance (Y) on a full logarithmic graph paper.
2.        Obtain the PAP value of patient by reference to the standard curve as follows: (These data are for demonstration purpose only and must not be used in place of data generated for each assay).
Well Description Absorbance PAP
No. (ng/ml) 450nm (ng/ml)
A1 0 0
B1 0
A2 1 0.043
B2 0.039
A3 3 0.139
B3 0.139
A4 5 0.24
B4 0.225
A5 15 0.88
B5 0.864
A6 30 1.686
B6 1.765
A7 PATIENT A 0.602 11.4
B7 0.591

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