Human p53 ELISA Assay

$560.00

The Human p53 ELISA Assay is to be used for the in-vitro quantitative determination of p53 in serum, plasma, or cell lysates. The assay will recognize both natural and recombinant p53. The kit recognizes p53 to different extents depending on the source of protein. The Eagle Biosciences Human p53 ELISA Assay kit is for Research Use Only.

SKU: P5331-K01 Categories: , ,

Human p53 ELISA Assay

The Human p53 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 1.5 U/ml
Dynamic Range: 3.1 U/mL – 100 U/ml
Incubation Time: 3h 45 min
Sample Type: Serum, Plasma, Cell Lysate
Sample Size: 100 µl


Specimen collection, processing & storage
Cell culture supernatants, serum, plasma or other biological samples will be suitable for use in the assay. Remove serum from the clot or red cells, respectively, as soon as possible after clotting and separation.
Cell culture supernatants:
Remove particulates and aggregates by spinning at approximately 1000 x g for 10 min.
Serum:
Avoid any unintentional stimulation of the cells by the procedure. Use pyrogen/endotoxin free collecting tubes. Serum should be removed rapidly and carefully from the red cells after clothing. For that, after clothing, centrifuge at approximately 1000 x g for 10 min and remove serum.
Plasma:
EDTA, citrate and heparin plasma can be assayed. Spin samples at 1000 x g for 30 min to remove particulates. Harvest plasma.
Storage:
If not analyzed shortly after collection, samples should be aliquoted (250-500μl) to avoid repeated freeze-thaw cycles and stored frozen at –70°C. Avoid multiple freeze-thaw cycles of frozen specimens.


Assay Principle


The p53 Kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for p53 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known p53 concentrations and unknowns are pipetted into these wells.

During the first incubation, the p53 antigen is added to wells.  After washing, a biotinylated monoclonal antibody specific for p53 is incubated. Then the enzyme (streptavidin-peroxydase) is added. After incubation and washing to remove all  unbound enzyme, a substrate solution which  acts on the bound enzyme is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of p53 present in the samples.


Related Products

Human CD21 ELISA Assay Kit
Human CD14 ELISA Assay Kit
Human CD116 ELISA Assay Kit

Additional Information

Assay Background


P53 (or TP53 for “tumor protein 53”) is a transcription factor regulating multiple important cellular functions such as cell cycle regulation, autophagy or apoptosis (programmed cell death). In humans, this gene is located on chromosome 172. The TP53 gene is inactivated in almost 50% of human cancers.

Assay Procedure

  1. Add 100 µl of  appropriate standard diluent to the blank wells
  2. Add 100 µl of sample to designated sample wells and 100 µl of the reconstituted control to the control wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 2 hours
  4. Remove the cover and wash the plate
  5. Add 50µl of diluted biotinylated anti-p53 to all wells
  6. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour
  7. Repeat wash step 5.
  8. Add 100μl of Streptavidin-HRP solution into all wells
  9. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  10. Repeat wash step 5.
  11. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  12. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  13. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve


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