High Sensitive IFN gamma ELISA Assay

Human CD138 ELISA Assay

$635.00

The Human CD138 ELISA Assay is a solid phase sandwich ELISA for the in-vitro qualitative and quantitative determination of sCD138 in supernatants, buffered solutions or serum and plasma samples. This assay will recognise both natural and recombinant human sCD138.  The Eagle Biosciences Human sCD138 ELISA Assay kit is for Research Use Only.

SKU: C3831-K01 Categories: , ,

Human CD138 ELISA Assay

The Human CD138 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 2.56 ng/ml
Dynamic Range: 8 ng/ml – 256 ng/ml
Incubation Time: 1h 45min
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: Syndecan-1

Assay Background

Syndecans are a transmembrane protein family within the heparin sulphate proteoglycan group that interact with many different molecules of the immune system through their heparin sulphate chains. The mammalian syndecan family consists of 4 proteins; syndecan 1 to 4 each encoded by very distinct genes. In adult tissues syndecan 1 (CD138) is predominantly expressed by epithelial cells and plasma cells (both normal and malignant) and currently considered the most reliable surface marker for plasma cells. In addition CD138 is also expressed on pre and immature B cells however this is regulated by IL-6 and LPS stimulation. Syndecan 1 has previously been shown to participate in cell to cell interactions, organ development, vessel formation and tissue regeneration following injury.

CD138 is regularly cleaved from the membrane and as a consequence high levels of soluble CD138 are found in the blood, which can be easily detected using a CD138 specific ELISA.

Via its heparin sulphate chains CD138 binds to and modulates the activity of a wide range of molecules involve in inflammation including chemokines, growth factors, selectins and other adhesion molecules. CD138 can also act as a receptor for collagen, fibronectin, thromobospondin and tenascin therefore involved in cell matrix adhesion. CD138 has been shown to mediate the binding of myeloma cells to type I collagen, and inhibits tumour cell invasion into collagen gels.

As CD138 has been shown to have important effects on tumour cell growth, survival, adhesion and invasion syndecan-1 may be an important regulator in cancer biology.

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Additional Information

Assay Principle

A capture Antibody highly specific for CD138 has been coated to the wells of the microtitre strip plate provided during manufacture. Binding of CD138 samples and known standards to the capture antibodies and subsequent binding of the biotinylated anti-CD138 secondary antibody to the analyte is completed during the same incubation period. Any excess unbound analyte and secondary antibody is removed. The HRP conjugate solution is then added to every well including the zero wells, following incubation excess conjugate is removed by careful washing. A chromogen substrate is added to the wells resulting in the progressive development of a blue coloured complex with the conjugate. The colour development is then stopped by the addition of acid turning the resultant final product yellow. The intensity of the produced coloured complex is directly proportional to the concentration of CD138 present in the samples and standards. The absorbance of the colour complex is then measured and the generated OD values for each standard are plotted against expected concentration forming a standard curve. This standard curve can then be used to accurately determine the concentration of CD138 in any sample tested.

  1. Add 100µl of each, Sample, Standard, Control and zero (appropriate standard diluent) in duplicate to appropriate number of wells
  2. Add 50µl of diluted biotinylated anti-CD138 to all wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour
  4. Remove the cover and wash the plate
  5. Add 100μl of Streptavidin-HRP solution into all wells
  6. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  7. Repeat wash step 4.
  8. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  9. Incubate in the dark for 12-15 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil.
  10. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve

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