Note: You clicked on an external link, which has been disabled in order to keep your shopping session open.
Western blot analysis of CREB (pS133) was performed by loading 20 µg of MDA-MB-231 (lane1), MCF7 (lane2), A549 (lane3), A431 (lane4) and HEK-293 (lane5) 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 1 hour at room temperature. CREB (ps133) was detected at ~43 kDa using ABfinity™ CREB Monoclonal Antibody (700129) at 1-3 µg/ml in 2.5 % skim milk at 4°C overnight on a rocking platform. To confirm specificity, competition was performed with phosphopeptide (10 µg/mL) as shown in the corresponding blot on right. 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|
|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 *|
|Flow Cytometry (Flow)||1-3µg/10^6 cells|
|Western Blot (WB)||1-3µg/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.
CREB (cAMP responsive element binding protein) encodes for a 43 kDa protein transcription factor and belongs to leucine zipper family. CREB binds to the cyclic-AMP response element (CRE, a sequence present in many viral and cellular promoters) as a homodimer. Phosphorylated form of CREB act as an inducer of transcription for the genes, upon hormone dependent stimulation of cyclic-AMP pathway. CREB is known to play important role in circadian rythm generation and memory formation; aberration in CREB-mediated gene expression has been linked with Alzheimer disease. In humans, the gene is located on the q arm of chromosome 2.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Active transcription factor CREB; cAMP responsive element binding protein 1; cAMP-response element-binding protein-1; cAMP-responsive element-binding protein 1; CREB-1; CREB1; cyclic adenosine 3',5'-monophosphate response element-binding protein CREB; Cyclic AMP-responsive element-binding protein 1; MGC9284; Transactivator protein
2310001E10Rik; 3526402H21Rik; AV083133; CREB; CREB-1; CREB1