HBEGF ELISA Kit

Catalog No. ABIN1672797

This Human HBEGF ELISA Kit is a Colorimetric ELISA Kit designed to quantify Human HBEGF.

Quick Overview for HBEGF ELISA Kit (ABIN1672797)

Target: HBEGF (Heparin-Binding EGF-Like Growth Factor (HBEGF))

Binding Specificity: AA 63-148

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.6-1000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Product Details HBEGF ELISA Kit

Minimum Detection Limit: 15.6 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human HBEGF

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: sf21
Immunogen sequence: D63-L148

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: sf21
Immunogen sequence: D63-L148

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Contains 1 EGF-like domain.

Plate: Pre-coated

Protocol: human HBEGF ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for HBEGF has been precoated onto 96-well plates. Standards(sf21, D63-L148) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for HBEGF is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human HBEGF amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL human HBEGF standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human HBEGF standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 92.7, Standard deviation: 5.10, CV(%): 5.5
• Sample 2: n=16, Mean(pg/ml): 369, Standard deviation: 16.97, CV(%): 4.6
• Sample 3: n=16, Mean(pg/ml): 621, Standard deviation: 23.6, CV(%): 3.8,
• Sample 1: n=24, Mean(pg/ml): 102, Standard deviation: 8.26, CV(%): 8.1
• Sample 2: n=24, Mean(pg/ml): 376, Standard deviation: 27.82, CV(%): 7.4
• Sample 3: n=24, Mean(pg/ml): 618, Standard deviation: 42.02, CV(%): 6.8

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

Target Details for HBEGF

Target: HBEGF (Heparin-Binding EGF-Like Growth Factor (HBEGF))

Alternative Name: HBEGF

Background:

Protein Function: Growth factor that mediates its effects via EGFR, ERBB2 and ERBB4. Required for normal cardiac valve formation and normal heart function. Promotes smooth muscle cell proliferation. May be involved in macrophage-mediated cellular proliferation. It is mitogenic for fibroblasts, but not endothelial cells. It is able to bind EGF receptor/EGFR with higher affinity than EGF itself and is a far more potent mitogen for smooth muscle cells than EGF. Also acts as a diphtheria toxin receptor.

Background: HBEGF(Heparin-binding EGF-like growth factor), also known as HEGFL or DTR, is a member of the EGF family of proteins that in humans is encoded by the HBEGF gene. The HBEGF gene is assigned to chromosome 5, thus confirming the assignment of the gene on the basis of its role in relation to diphtheria toxin susceptibility. HB-EGF is an 87 amino acid glycoprotein which displays highly regulated gene expression. It has been shown to play a role in wound healing, cardiac hypertrophy and heart development and function. HB-EGF binding and activation of EGF receptors plays a critical role during cardiac valve tissue development and the maintenance of normal heart function in adults.

Synonyms: Proheparin-binding EGF-like growth factor,Heparin-binding EGF-like growth factor,HB-EGF,HBEGF,Diphtheria toxin receptor,DT-R,HBEGF,DTR, DTS, HEGFL,

Full Gene Name: Proheparin-binding EGF-like growth factor

Cellular Localisation: Heparin-binding EGF-like growth factor: Secreted, extracellular space. Mature HB-EGF is released into the extracellular space and probably binds to a receptor.

Gene ID: 1839

UniProt: Q99075

Pathways: RTK Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway