Human CD44 ELISA Assay

$560.00

The Human CD44 ELISA Assay is to be used for the in-vitro quantitative determination of soluble CD44 (CD44/phagocytic glycoprotein-1/Pgp-1) in human serum, plasma, buffered solutions or cell culture medium. The assay will recognize both natural and recombinant human CD44. The Eagle Biosciences Human CD44 ELISA Assay kit is for Research Use Only.

SKU: C4431-K01 Categories: , ,

Human CD44 ELISA Assay

The Human CD44 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.13ng /ml
Dynamic Range: 0.31 ng/ml – 10 ng/ml
Incubation Time: 2h 10 min
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: Phagocytic Glycoprotein-1, Pgp-1


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 CD44 Kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for CD44 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known CD44 concentrations and unknowns are pipetted into these wells.  During the first incubation, the CD44 antigen, standards and zeros are bound this is followed by a separate incubation where the biotinylated monoclonal antibody specific for CD44 is added.  After washing, the enzyme (streptavidin-peroxydase) is added. After incubation and washing to remove all the unbound enzyme, a substrate solution is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of CD44 present in the samples.


Related Products

Human CD54 ELISA Assay
Human CD31 ELISA Assay Kit
Human CD86 ELISA Assay Kit

Additional Information

Assay Background


P glycoprotein, also called P-GP, “Permeability-GlycoProtein” or CD44, is a glycoprotein causing a phenomenon of multi-drug resistance.

Discovered in the 1980s, the existence of CD44 was revealed during experiments on the causes of resistance to chemotherapeutic agents of certain tumors.

Assay Procedure

  1. Add 100µl of each standard, sample, control and zero (Standard Diluent) in duplicate to appropriate number of wells
  2. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour
  3. Remove the cover and wash the plate
  4. Add 50µl of diluted biotinylated anti-CD44 to all wells
  5. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 mins
  6. Repeat wash step 3.
  7. Add 100μl of Streptavidin-HRP solution into all wells
  8. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  9. Repeat wash step 3.
  10. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  11. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  12. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve


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