Description
Transcription of various genes is regulated by the cyclic AMP (cAMP) signal transduction pathway through a family of cAMP Responsive Element (CRE)-Binding transcription factors that include CREB, CREM (CRE Modulator), and ATF-1 (Activating Transcription Factor 1). The genes for these transcription factors encode multiple isoforms that are created by alternative splicing, alternative initiation codons, and alternative intronic promoters. Homo- and heterodimers of these members of the basic domain-and-leucine zipper superfamily of proteins bind the palindromic TGACGTCA sequence of CRE. CREB is expressed in all somatic cells, and many different stimuli activate CREB by phosphorylation of its serine 133 (S133). The transcriptional activity and specificity of CREB are regulated by the signaling pathways initiated by the various stimuli. Transcriptional regulation by CREB family members has been implicated in many physiological responses to extracellular and environmental stimuli, such as memory, tissue growth and development, and homeostasis. The J151-21 monoclonal antibody recognizes the phosphorylated S133 of human CREB in addition to cross-reacting with S63 of human ATF-1.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
5. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
6. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.