Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 mins (after plate preparation)
Sample Type & Volume Required
Cell lysates (100 µL)
Range
125.00 - 8,000 pg/mL
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Specificity
Please see the
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
Product Features
Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
Development protocols are provided to guide further assay optimization
Assay can be customized to your specific needs
Available in 2, 5, and 15- (96-well) plate pack sizes
Economical alternative to Western blot
Kit Content
Capture Antibody
Conjugated Detection Antibody
Calibrated Immunoassay Standard or Control
Streptavidin-HRP
Other Reagents Required
PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na 2HPO 4, 1.5 mM KH 2O 4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # ), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
Substrate Solution: 1:1 mixture of Color Reagent A (H 2O 2) and Color Reagent B (Tetramethylbenzidine) (Catalog # )
Stop Solution: 2 N H 2SO 4 (Catalog # )
Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # ), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # ), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Data Examples
Figure 1: The Human Total ErbB4 DuoSet IC ELISA is more sensitive than Western blot analysis. Lysates prepared from MDA-MB-453 human breast cancer cells were serially diluted and analyzed by (A) Western Blot and (B) this DuoSet® IC ELISA. Immunoblots were incubated with an anti-ErbB4 polyclonal antibody to detect total ErbB4. Bands were visualized with anti-rabbit IgG-HRP (Catalog # ) followed by chemiluminescent detection. Human ErbB4 can be detected in this DuoSet® IC ELISA by using approximately 5 to 10 times less lysate than is needed for a conventional IP-Western Blot. |
Figure 2: The Human Total ErbB4 DuoSet IC ELISA measures the relative level of ErbB4. Lysates were prepared from MDA-MB-453 human breast cancer cells and the MCF-7 human breast cancer cell line. Both ELISA and Western Blot (inset) analyses were performed using 100 g of lysate. The Western Blot was performed as described in Figure 1. |
Preparation and Storage
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: ErbB4/Her4
ErbB4, also known as Her4, is a type I membrane glycoprotein that is a member of the ErbB family of tyrosine kinase receptors. ErbB family members serve as receptors for the EGF family of growth factors. ErbB4 ligands include the neuregulins, betacellulin and heparin-binding EGF-like growth factor (HB-EGF).
Entrez Gene IDs:
2066 (Human); 13869 (Mouse); 59323 (Rat);
Long Name:
Receptor Tyrosine Protein Kinase ErbB4
Aliases:
avian erythroblastic leukemia viral (v-erb-b2) oncogene homolog 4; EC 2.7.10; EC 2.7.10.1; ErbB4; HER4; HER4MGC138404; p180erbB4; Proto-oncogene-like protein c-ErbB-4; receptor tyrosine-protein kinase erbB-4; Tyrosine kinase-type cell surface receptor HER4; v-erb-a avian erythroblastic leukemia viral oncogene homolog-like 4; v-erb-a erythroblastic leukemia viral oncogene homolog 4 (avian)
