Human Phospho-PDGF R alpha DuoSet IC ELISA Summary

Product Summary

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated human PDGF R alpha in cell lysates. An immobilized capture antibody specific for PDGF R alpha binds both phosphorylated and unphosphorylated PDGF R alpha. After washing away unbound material, an HRP-conjugated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Preparation and Storage

Background: PDGF R alpha

PDGF R alpha (Platelet-Derived Growth Factor Receptor alpha) is a transmembrane receptor tyrosine kinase that can form homodimers or heterodimers with PDGF R beta when engaged by dimers of the PDGF ligands. PDGF R alpha binds to PDGF-AA, PDGF-AB, and PDGF-CC. It is strongly expressed in oligodendrocyte, lung, skin, and intestinal progenitor cells, at low levels in most mesenchymal cells, and can be induced by inflammation. PDGF R alpha function is important during formation of the cranial and cardiac neural crest, retina, gonads, lung alveoli, intestinal villi, skin, hair follicles, skeleton, teeth, palate, and interstitial kidney mesenchyme. Dysregulated PDGF R alpha signaling can result in spina bifida or cleft palate-type malformations. Postnatally, PDGF R alpha is implicated in gliomas and fibrotic disorders of lung, heart, and skin (scleroderma).

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.