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Human HB-EGF

Human HB-EGF Quantikine ELISA Kit Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

4.5   hours

Sample   Type & Volume Required Per Well

Cell   Culture Supernates (50 uL), Serum (100 uL), EDTA Plasma (100 uL),   Platelet-poor EDTA Plasma (100 uL), Heparin Plasma (100 uL), Platelet-poor   Heparin Plasma (100 uL), Saliva (50 uL), Urine (100 uL), Human Milk (100 uL)

Sensitivity

1.74   pg/mL

Assay   Range

7.8 -   500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Platelet-poor EDTA   Plasma, Heparin Plasma, Platelet-poor Heparin Plasma, Saliva, Urine, Human   Milk)

Specificity

Natural   and recombinant human HB-EGF

 

Cross-reactivity

<   0.5% cross-reactivity observed with available related molecules.< 50%   cross-species reactivity observed with species tested

Interference

No   significant interference observed with available related molecules.

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

Shipping

The   product is shipped at ambient temperature. Upon receipt, store it immediately   at the temperature recommended below.

Storage

Store   the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: HB-EGF

HB-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.

Long   Name:

Heparin   Binding EGF-like Growth Factor

Entrez   Gene IDs:

1839   (Human); 15200 (Mouse)

Alternate   Names:

diphtheria   toxin receptor (heparin-binding epidermal growth factor-like growthfactor);   Dtr; DTRHEGFLdiphtheria toxin receptor (heparin-binding EGF-like growth   factor); Dts; DTSF; HBEGF; HB-EGF; Hegfl; heparin-binding EGF-like growth   factor; heparin-binding epidermal growth factor; proheparin-binding EGF-like   growth factor

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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