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Prostate Specific Antigen ELISA Kit (PSA) ELISA Kit

PSA Reactivity: Human Colorimetric Sandwich ELISA Plasma
Catalog No. ABIN5067601
$742.50
Plus shipping costs $45.00
96 tests ABIN5067601
96 tests ABIN5067601
local_shipping Shipping to: United States
Delivery in 2 to 3 Business Days
  • Target
    Reactivity
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Application
    ELISA
    Sample Type
    Plasma, Serum
    Analytical Method
    Quantitative
    Characteristics
    Free PSA ELISA Kit is an enzyme immunoassay developed for detection and quantitation of the human f-PSA protein. The kit has a detection sensitivity limit of 15 pg/mL and contains a HYBRITECH® f-PSA Calibration Standard. Each kit provides sufficient reagents to perform up to 96 assays including standard curve and f-PSA samples. Note: This kit is specific for f-PSA and does not detect bound PSA.
    Components
    1. Anti-f-PSA Antibody Coated Plate : One strip well 96-well plate.
    2. Biotinylated Anti-f-PSA Antibody (1000X) : One 20 μL vial.
    3. Streptavidin-Enzyme Conjugate : One 20 μL vial.
    4. Assay Diluent : One 50 mL bottle.
    5. 10X Wash Buffer : One 100 mL bottle.
    6. Substrate Solution : One 12 mL amber bottle.
    7. Stop Solution (Part. No. 310808): One 12 mL bottle.

    Box 2 (shipped on blue ice packs)

  • Application Notes
    Optimal working dilution should be determined by the investigator.
    Plate
    Pre-coated
    Protocol
    An anti-f-PSA coating antibody is adsorbed onto a microtiter plate. Free PSA protein present in the sample or standard binds to the antibodies adsorbed on the plate, a biotinylated anti-f-PSA antibody is added and binds to the antigen captured by the first antibody. Following incubation and wash steps, a streptavidin-enzyme conjugate is added and binds to the biotinylated anti-f-PSA antibody. Unbound streptavidin-enzyme conjugate is removed during a wash step, and substrate solution is added to the wells. A colored product is formed in proportion to the amount of f-PSA present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450 nm. A standard curve is prepared from a f-PSA calibrator and sample concentration is then determined.
    Reagent Preparation
    • 1X Wash Buffer: Dilute the 10X Wash Buffer to 1X with deionized water. Stir to homogeneity.
    • Biotinylated Anti-f-PSA Antibody and Streptavidin-Enzyme Conjugate: Immediately before use dilute the Biotinylated Anti-f-PSA Antibody 1:1000 and the Streptavidin-Enzyme Conjugate 1:1000 with Assay Diluent. Do not store diluted solutions. 3 Preparation of Standard Curve 1. Prepare a dilution series of f-PSA Standard in the concentration range of 1 ng/mL - 0.016 ng/mL by diluting the stock solution in Assay Diluent (Table 1). Standard 10 ng/mL HYBRITECH® A s s a y Diluent HYBRITECH® Tubes f-PSA Standard (μL) (μL) f-PSA (ng/mL) 1 80 720 1 2 400 of Tube #1 400 0.5 3 400 of Tube #2 400 0.25 4 400 of Tube #3 400 0.125 5 400 of Tube #4 400 0.063 6 400 of Tube #5 400 0.031 7 400 of Tube #6 400 0.016 8 0 400 0 Table 1.
    Assay Procedure
    1. Prepare and mix all reagents thoroughly before use.
    2. Add 100 μL of PSA sample or standard to the Anti-t-PSA Antibody Coated Plate. Each PSA sample, standard, blank, and control should be assayed in duplicate.
    3. Cover with a plate cover and incubate at room temperature for 1 hour on an orbital shaker.
    4. Remove plate cover and empty wells. Wash microwell strips 5 times with 250 μL 1X Wash Buffer per well with thorough aspiration between each wash. After the last wash, empty wells and tap microwell strips on absorbent pad or paper towel to remove excess 1X Wash Buffer.
    5. Add 100 μL of the diluted Biotinylated Anti-t-PSA Antibody to each well.
    6. Cover with a plate cover and incubate at room temperature for 1 hour on an orbital shaker.
    7. Remove plate cover and empty wells. Wash the strip wells 5 times according to step 4 above.
    8. Add 100 μL of the diluted Streptavidin-Enzyme Conjugate to each well.
    9. Cover with a plate cover and incubate at room temperature for 1 hour on an orbital shaker.
    10. Remove plate cover and empty wells. Wash microwell strips 5 times according to step 4 above. Proceed immediately to the next step. 10
    11. Warm Substrate Solution to room temperature. Add 100 μL of Substrate Solution to each well, including the blank wells. Incubate at room temperature on an orbital shaker. Actual incubation time may vary from 5-20 minutes. Note: Watch plate carefully, if color changes rapidly, the reaction may need to be stopped sooner to prevent saturation.
    12. Stop the enzyme reaction by adding 100 μL of Stop Solution into each well, including the blank wells. Results should be read immediately (color will fade over time).
    13. Read absorbance of each microwell on a spectrophotometer using 450 nm as the primary wave length.
    Restrictions
    For Research Use only
  • Storage
    4 °C
    Storage Comment
    Upon receipt, store all components at 4°C.
  • Target
    Alternative Name
    PSA (PSA ELISA Kit Abstract)
    Background
    Prostate-Specific Antigen (PSA) is a 33 kDa serine proteinase secreted by epithelial cells of the prostate gland. PSA exists in serum in 3 main forms: unbound free PSA (f-PSA), α1-antichymotrypsin bound PSA, and α2-macroglobulin bound PSA. Blood PSA levels are typically low in men with healthy prostates, but are often elevated in individuals with prostate disorders (i.e. cancer, prostatitis, benign prostate hyperplasia). Measuring the ratio of free to total PSA (t-PSA) is an effective tool in prostate cancer screening and diagnosis.
    Pathways
    Complement System
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