Humanized Anti-HER-2 (Herceptin/Trastuzumab) ELISA Assay Kit

$725.00

The Eagle Biosciences Humanized Anti-HER-2 (Herceptin/Trastuzumab) ELISA Assay kit is intended for use in the quantitative determination of Herceptin levels in human serum and EDTA plasma. The Humanized Anti-HER-2 (Herceptin/Trastuzumab) ELISA Assay kit is intended for research use only.

Humanized Anti-HER-2 (Herceptin/Trastuzumab) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.245 ng/mL
Dynamic Range: 0.245 – 500 ng/mL
Incubation Time: >2 hours
Sample Type: Serum or EDTA Plasma
Sample Size: 10 µL

Product Developed and Manufactured in the USA


Additional Information

Assay Background

Herceptin (Trasuzumab) is a humanized monoclonal antibody used in cancer treatment for Human Epidermal Growth Factor Receptor 2 (HER2) over-expressing tumors. In the case of these tumors, Herceptin binds to and inhibits tumor growth. Herceptin can also be labeled with various toxins to enhance tumor suppression. Studies have also found that Herceptin targets gastric cancer stem cells characterized by CD90 phenotype.

Assay Principle

The Eagle Biosciences Humanized Anti-HER-2 (Herceptin/Trastuzumab) ELISA Assay kit utilizes the “sandwich” technique with HER2 coated to solid phase microtiter plate wells and an antibody to human IgG which is labeled with horseradish peroxidase used for detection. Assay calibrators, controls and diluted patient samples are added directly to wells of a micro titer plate that is coated with HER2 recombinant protein. Subsequently, horseradish peroxidase (HRP) conjugated human IgG antibody is added to each well. After the first incubation period, a “sandwich” of “HER2 – Herceptin – HRP conjugated human IgG antibody” is formed on the surface of the plate wells. The unbound proteins are removed in the subsequent washing step. For the detection of this immunocomplex, the wells are then 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 wall of each micro titer well is directly proportional to the amount of Herceptin in the calibrators. A calibration curve is generated by plotting the absorbance versus the respective Herceptin concentration for each calibrator using point-to-point or 4 parameter curve fitting. The concentration of Herceptin in test samples is determined directly from this calibration curve.

Assay Procedure

  1. Add 25 µL of calibrators, controls and diluted 1:100 test samples into the designated microwells.
  2. Immediately add 100 µL of working 1x assay buffer into the designated microwells.
  3. Seal the plate wells securely, cover with foil or other material to protect from light and rotate on an ELISA plate shaker (small orbit radius) for 1 hour at 400 to 450 rpm.
  4. 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 Herceptin Tracer Antibody to each well.
  6. Seal the plate wells securely, cover with foil or other material to protect from light and rotate for 30 minutes on an ELISA plate shaker (small orbit radius) at 400 to 450 rpm.
  7. 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 aluminum foil or other material to avoid exposure to light. Incubate plastic static, at room temperature for 20 minutes.
  10. Immediately add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
  11. Read the absorbency at 450 nm.

Typical Standard Curve

Manual

Product Manual