Western blot analysis of STAT6 was performed by loading 20 µg of HeLa (lane1), A431 (lane2) and A549 (lane3) cell lysates using Novex® NuPAGE® 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. STAT6 was detected at ~100 kDa using ABfinity™ STAT6 Recombinant Rabbit Oligoclonal Antibody (710140) at 0.5 µg-1 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 630–638 of human STAT6|
|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)||3µg|
|Western Blot (WB)||0.5-1µg/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™ 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.
STAT6 (Signal Transducers and Activators of Transcription 6) belongs to the STAT protein family, which in response to cytokines and growth factors are phosphorylated by receptor-associated kinases, and act as transcription factors. When induced by CD80 or CD86 ligand binding, phosphorylated CD28 recruits and activates STAT6 which in turn translocates to the nucleus and binds to a STAT6-responsive element on the human IL-4 promoter. STAT6 is crucial for IL-4-mediated biological responses, and is also involved in processes mediated by IL-13, a cytokine related to IL-4. IL4 and IL13 have been shown to induce the production of IgE, a major mediator in an allergic response. STAT6 occurs naturally in two isoforms: STAT6a and STAT6c
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.