Cancer Biomarker ELISA Assay

Chromogranin A ELISA Kit

$900.00

The Chromogranin A ELISA Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of human Chromogranin A levels in EDTA-plasma and serum samples. This assay exclusively measures human Chromogranin A without the high dose “hook” effect up to 1,000,000 ng/ml. The Eagle Biosciences Human Chromogranin A ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: CHR31-K01 Categories: , ,

Chromogranin A ELISA Kit

Chromogranin A ELISA Kit Developed and Manufactured in the USA

Size: 1×96 wells
Sensitivity: 2 ng/mL
Dynamic Range: 19.2 – 660 ng/ml
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma
Sample Size: 15 µL
Alternative Names: Human Chromogranin A ELISA Kit, CgA ELISA
For Research Use Only

Controls Included

Assay Principle

The Chromogranin A ELISA Kit is designed, developed and produced for the quantitative measurement of human chromogranin A in EDTA-plasma or serum sample. The assay utilizes the two-site “sandwich” technique with two selected antibodies that bind to different epitopes of human Chromogranin A.

Assay standards, controls and samples are added directly to wells of microplate that is coated with a polyclonal chromogranin A antibody. After the first incubation period, the antibody on the wall of microtiter well captures human chromogranin A in the sample and unbound proteins in each microtiter well is washed away. Then a horseradish peroxidase (HRP) labeled monoclonal anti-human chromogranin A antibody is added to each microtiter well and a “sandwich” of “monoclonal antibody – human chromogranin A – polyclonal antibody” is formed. The unbound monoclonal antibody is removed in the subsequent washing step. For the detection of this immunocomplex, the well is incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the chromogranin A on the wall of the microtiter well is directly proportional to the amount of chromogranin A in the sample. A standard curve is generated by plotting the absorbance versus the respective human chromogranin A concentration for each standard with a 4 parameter curve fit. The concentration of human chromogranin A in test samples is determined directly from this standard curve.

Place a sufficient number of antibody coated microwell strips in a holder to run human chromogranin A standards, controls and unknown samples in duplicate.

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Additional Information

Test Configuration for Chromogranin A ELISA Assay Kit:

ROW STRIP 1 STRIP 2 STRIP 3
A STD 1 STD 5 SAMPLE 2
B STD 1 STD 5 SAMPLE 2
C STD 2 C 1 SAMPLE 3
D STD 2 C 1 SAMPLE 3
E STD 3 C 2 SAMPLE 4
F STD 3 C 2 SAMPLE 4
G STD 4 SAMPLE 1
H STD 4 SAMPLE 1

Assay Procedure

  1. Add 15 µL of standards, controls and patient samples into the designated microwells.
  2. Add 200 µL of assay buffer to each well
  3. Cover the plate with one plate sealer and incubate plate on an ELISA plate shaker with a shaking rate at 350 rpm to 450 rpm at room temperature for 1 hour.
  4. Remove plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  5. Add 100 µL of Chromogranin A Tracer Antibody to each of the wells.
  6. Cover the plate with the plate sealer and incubate plate on an ELISA plate shaker with a shaking rate at 350 rpm to 450 rpm at room temperature for 1 hour.
  7. Remove plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
  8. Add 100 µL of ELISA HRP Substrate into each of the wells.
  9. Cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
  10. Incubate plate at room temperature for 20 minutes.
  11. Remove the aluminum foil and plate sealer. Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  12. Remove the aluminum foil and plate sealer. Add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  13. Read the absorbance at dual wave length at 450/650 nm or 450/620 nm within 10 minutes in a microplate reader and select a 4-parameter curve fit for result calculation.

Typical Standard Curve

chromogranin-a-elisa-kit

 

Package Inserts

 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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