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Human Prostate Specific Antigen (PSA)

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Human Prostate Specific Antigen (PSA) General References 1. Wang MC (1979) Invest. Urol. 17, 159; (2) Frankel AE 91982) Cancer Res. 42, 3714. Instruction Manual No. M-EK-1500 (3) Papsider LD (1980) Cancer Res. 40, 2428. (4) Ponts JE (1982) J. Urol. 128, 1216. (5) Killan CS (1985) Cancer Res. 45, 886. (6) Kuriyam M (1980) Cancer Res. 40, 4658. (7) Kuriyam M (1982) J Natl. Cancer 68, 99. (8) Schiffman RB (1987) Clin Chem. 33, 2086 Related ELISA kits for other serum proteins CatalogNumber ItemName EK-0050 Human Neuron Specific Enolase (NSE) ELISA Kit Human Prostate Specific Antigen (PSA) EK-1750 Human IgG (total) ELISA Kit EK-1760 Human IgM ELISA Kit EK-1800 Human IgE ELISA Kit EK-0410 Human Chorionic Gonadotropin (HCG-free beta) ELISA Kit EK-0040 Human C-peptide ELISA Kit EK-0200 Human Folicle Stimulating Hormone (FSH) ELISA Kit ELISA KIT Cat. No. EK-1500 EK-0100 Human Luteinizing Hormone (LH) ELISA Kit EK-0030 Human Insulin ELISA Kit EK-0300 Human Prolactin (PRL) ELISA Kit EK-0400 Human Chorionic Gonadotropin (HCG) ELISA Kit EK-0500 Human Alpha Fetoprotein (AFP) ELISA Kit EK-1000 Human C-Reactive Protein (CRP) ELISA Kit For Quantitative Determination of Human PSA In Serum EK-1200 Human Albumin ELISA Kit EK-0020 Human Beta-2 microglobulin (B2M) ELISA Kit EK-1810 Human Ferritin ELISA Kit EK-0700 Human Sex Hormone Binding Glob (SHBG) ELISA Kit EK-1210 Human Transferrin (Tf) ELISA Kit EK-0010 Human Leptin ELISA Kit EK-1190 Human Serum Albumin ELISA Kit EK-1840 Human Pancreatic & GI Cancer (CA199) ELISA Kit EK-1830 Human Ovarian Cancer (CA153) ELISA Kit EK-1310 Human Pancreatic Lipase ELISA Kit EK-1600 Human Growth Hormone (GH) ELISA Kit EK-1510 Human Free Prostate Specific Antigen (PSA-free) ELISA Kit EK-1500 Human Prostate Specific Antigen (PSA) ELISA Kit EK-1400 Human Prostatic Acid Phosphatase (PAP) ELISA Kit EK-0900 Human IGF-Binding Protein 1 (IGFBP1) ELISA Kit EK-0060 Human Pancreatic Colorectal cancer (CA-242) ELISA Kit EK-1820 Human Ovarian Cancer (CA125) ELISA Kit Page 7 (Rev. ub61025A) Human Prostate Specific Antigen (PSA) ELISA KIT # EK-1500 PERFORMANCE CHARACTERISTICS For Quantitative Determination of Human PSA In Serum 1. DETECTION LIMIT Kit Contents: (reagents for 96 tests) Based on twenty replicate determinations of the zero standard, the minimum concentration of human PSA detected using this assay is 0.3 ng/ml. The C o m p o n e n t s C a t # detection limit is defined as the value deviating by 2 SD from the zero standard. Anti-human PSA coated microwell strip plate 1 5 0 1 2. PRECISION (96 wells), Ready-to-use PSA Standard A (ready-to-use), 11 ml 1 5 0 2 Intra-assay precision: PSA Standard B (ready-to-use), 0.75 ml 1 5 0 3 Three serum samples were run in ten replicates in an assay. The samples PSA Standard C (ready-to-use), 0. 75 ml 1 5 0 4 showed good intra-assay precision (7.8-12 %CV). The actual values were: mean 2.31 ng/ml, SD 0.27 ng/ml, %CV 11.7; mean 23.92 ng/ml, SD 2.11 ng/ml, %CV PSA Standard D (ready-to-use), 0. 75 ml 1 5 0 5 8.8; mean 45.1 ng/ml, SD 3.4 ng/ml, %CV 7.8, respectively PSA Standard E (ready-to-use), 0. 75 ml 1 5 0 6 Inter-assay precision: PSA Standard F (ready-to-use), 0. 75 ml 1 5 0 7 Three serum samples were run in duplicate in eight independent assays. The Anti-human PSA IgG-HRP Conjugate; 11 ml 1 5 0 8 samples showed good inter-assay precision (7-10 %CV). The actual values were: mean 2.8 ng/ml, SD 0.23 ng/ml, %CV 8.2; mean 25.1 ng/ml, SD 2.44 Wash buffer (20X), 50 ml W B - 2 0 ng/ml, %CV 9.7; mean 47.1 ng/ml, SD 3.4 ng/ml, %CV 7.3, respectively. HRP substrate Buffer; Solution A; 11 ml 1 5 0 0 S A 3. RECOVERY HRP substrate, Solution B; 11 ml 1 5 0 0 S B A known amount of hPSA (35, and 76 ng/ml) was added to three patient sera Stop solution (ready-to-use), 10 ml T - 1 0 (with original PSA concentrations of 6.2 and 44 ng/ml) and the total PSA concentrations measured. The assay showed excellent mean recoveries of Complete Instruction Manual M - E K - about 95% (range 92-105%). 1 5 0 0 4. SPECIFICITY The specificity of PSA kit was determined by measuring interference from high Introduction concentrations of human serum proteins, hormones, and tumor markers. No cross reactivity was observed. The human PSA ELISA kit #1500 detects total Human Prostate-specific antigen (PSA) is a single chain glycoprotein with PSA. GSI has another ELISA kit #1510 for free PSA. Mol. Wt. Of 34 kDa containing 7% carbohydrates. It is uniquely associated with prostate tissue and presents in both normal and cancerous prostatic Species crossreactivity and use of other biological fluids tissue, prostatic fluid and seminal plasma (1). PSA is not present in any of the normal tissue obtained from men nor is it produced by cancers of the Human PSA kit has not been tested in other species (mouse, rat, monkey etc). lung, colon, rectum, stomach, breast, pancreas or thyroid (2). The kit may work in monkey samples but the reactivity witt other animals is less likely. Elevated serum PSA concentration have been reported in patient with prostate cancer, benign prostatic hypertrophy, or inflammatory tissues, but Human PSA kit has been validated for use in serum. However, it has been used not in apparently healthy women with cancer (3). PSA levels correlate with to detect PSA released into culture medim by some cancer cells (LnCap etc). It the stage of disease and response to treatment in-patients with prostatic is recommended to include fresh culture media to establish baseline values or cancer (4-7). Therefore sequential measurements of PSA concentration any crossreactivity due to serum components. See published references on can be an important tool in monitoring patients with prostatic cancer and in page 7. determining the potential and effectiveness of surgery of other therapies. GSI’s PSA ELISA kit provides for the measurement of PSA in serum for Page 6 monitoring patients with prostatic cancer. Page 1 WORKSHEET OF TYPICAL ASSAY PRINCIPLE OF THE TEST Mean Net Calculated PSA ELISA kit is based on simultaneous binding of human prostate specific Wells Stds/samples A450 nm A450 nm Conc. antigen (PSA) from samples to two antibodies, one immobilized on microtiter well (ng/ml) (ng/ml) plates, and other conjugated to the enzyme horseradish peroxidase. After a A1, A2 Std. A (0) 0.00 washing step, chromogenic substrate is added and colors developed. The enzymatic reaction (color) is directly proportional to the amount of PSA present in B1, B2 Std. B (1.5) 0.040 0.040 the sample. Adding stopping solution terminates the reaction. Absorbance is C1, C2 Std. C (5) 0.143 0.143 then measured on a microtiter well ELISA reader at 450 nm. and the D1, D2 Std. D (15) 0.435 0.435 concentration of PSA in samples and control is read off the standard curve. E1, E2 Std. E (30) 0.865 0.865 F1, F2 Std. F (60) 1.706 1.706 MATERIALS AND EQUIPMENT REQUIRED G1, G2 Sample 1 0.400 0.400 14.5 Adjustable micropipet (10-200 ul) and multichannel pipet with disposable plastic tips, Reagent troughs, plate washer (recommended) and ELISA plate Reader. NOTE: These data are for demonstration purpose only. A complete standard curve must be run in every assay to determine sample values. PRECAUTIONS Each laboratory should determine their own normal reference values. The Genemed Synthesis’s PSA ELISA test is intended for in vitro research use only. The reagents contain thimerosal as preservative; necessary care should be taken when disposing solutions. The Control Serum has been prepared from human sera shown to be negative for HBsAg and HIV antibodies. Nevertheless, such tests are unable to prove the complete absence of viruses, therefore, sera should be handled with appropriate precautions. Applicable MSDS, if not already on file, for the following reagents can be obtained from GSI or the web site. TMB (substrate), H2SO4 (stop solution), and Prolcin-300 (0.1% v/v in standards, sample diluent and HRP-conjugates). SPECIMEN COLLECTION AND HANDLING Collect blood by venipuncture, allow to clot, and separate the serum by centrifugation at room temperature. Do not heat inactivate the serum.. If sera can not be immediately assayed , these could be stored at -20oC for up to six months. Avoid repeated freezing and thawing of samples. No preservatives should be added to the serum. Stability of Samples PSA is unstable at 4oC and 20oC. Samples should not be stored at room temp or 4oC for more than 24 hours. It is recommended to freeze all samples as soon as possible. Avoid repeated freeze and thaw. Preparation of the reagents A typical standard assay curve (do not use this for calculating sample 1. Dilute the wash buffer 1:20 with water. Diluted buffer can be kept at values) 4oC for 6-12 months. Dilute 25 ml of 20X stock in 1000 ml water. 2. Prepare working solution of TMB substrate by mixing equal volume of solution of A and B. Prepare only in required amounts. Do not store mixed solutions of A and B for more than 30 mins.
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