Western blot analysis of ERBB2 was performed by loading 30 µg of MCF7 (lane1),HEK-293 (lane2), Hep G2 (lane3) and SK-Br-3 (lane4) cell lysate using NuPAGE® Novex® 4-12% Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800). Proteins were transferred to a PVDF membrane and blocked with 5% skim milk for 1 hour at room temperature. ERBB2 was detected at ~138 kDa using ABfinity™ ERBB2 Recombinant Rabbit Monoclonal Antibody (701399) at 1-2 µg/mL in 2.5% skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 33–46 of human ERBB2|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1-2 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
ERBB2 is a member of the epidermal growth factor receptor family of tyrosine kinases. Unlike other receptors, ERBB2 does not have a ligand binding domain, and acts by binding other ligand bound EGF receptors. This heterodimer formation results in mediation of various signaling pathways and downstream effectors. In humans, the gene encoding ERBB2 is present on chromosome 17.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.