Western blot analysis of p44/42 MAPK (ERK1/2) [pT202/pY204] was performed by loading 20 µg of NIH/3T3 (lane1) and NIH/3T3 treated for 10 minutes with 100 nM/ml of EGF (lane2) cell lysates using Novex® NuPAGE® 4-12% Bis-Tris gel (NP0321BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for overnight at 4°C. p44/42 MAPK (ERK1/2) [pT202/pY204] was detected at ~42 kDa using ABfinity™ p44/42 MAPK (ERK1/2) [pT202/pY204] Recombinant Rabbit Oligoclonal Antibody (710318) at 0.5-1 µg/mL in 2.5% skim milk at room temperature for 3 hours 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, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosopeptide corresponding to amino acids 197–209 of human MAPK [pT202/pY204]|
|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 *|
|ChIP assay (ChIP)||1 ul|
|Western Blot (WB)||0.5-2 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.
This antibody is predicted to react with mouse, rat, non-human primate and rabbit based on sequence homology.
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™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
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.
Mitogen activated protein kinase (MAPK) belongs to a family of serine/threonine kinases that are activated upon exposure of cells to a variety of stimuli including mitogens, hormones, growth factors, cytokines and bioactive peptides. There are two closely related isoforms of MAPK, with apparent molecular weights of 44 kDa and 42 kDa. They are abundantly expressed in neurons of the mature central nervous system. Activated MAPK plays a role in cell division and differentiation, and regulation of the cytoskeleton.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.