Human HB-EGF | ||||||||||||||||||||||||||||
Human HB-EGF Quantikine ELISA Kit Summary
Product Summary The Quantikine Human HB-EGF Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human HB-EGF in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains Sf 21-expressed recombinant human HB-EGF and antibodies raised against the recombinant factor. Results obtained using natural human HB-EGF showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human HB-EGF. Preparation and Storage
Background: HB-EGFHB-EGF (Heparin-Binding EGF-like growth factor), also known as DTR, is produced by bronchial epithelium, visceral and vascular smooth muscle, CD4+ T cells, cardiac muscle, glomerular podocytes, keratinocytes, and IL-10-secreting regulatory macrophages. It activates both the EGF R/ErbB1 and ErbB4 receptors and has been linked to many cellular processes including proliferation, apoptosis, cell migration/invasion, differentiation, morphogenesis, and development. HB-EGF is also involved in several aspects of cancer development and progression.
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. |
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