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Alexa Fluor® 647 Mouse Anti-CREB (pS133) / ATF-1 (pS63)
Alexa Fluor® 647 Mouse Anti-CREB (pS133) / ATF-1 (pS63)

Analysis of CREB (pS133) / ATF-1 (pS63) in human peripheral blood lymphocytes.  Human whole blood was either stimulated with PMA and Ionomycin for 15 minutes at 37ºC (shaded histogram) or unstimulated (open histogram).  The cells were lysed and fixed with 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10-15 minutes at 37ºC, then permeabilized usingBD Phosflow™ Perm Buffer II (Cat. No. 558052) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-CREB (pS133) / ATF-1 (pS63) antibody.  For data analysis, lymphocytes were selected by scatter profile.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Analysis of CREB (pS133) / ATF-1 (pS63) in human peripheral blood lymphocytes.  Human whole blood was either stimulated with PMA and Ionomycin for 15 minutes at 37ºC (shaded histogram) or unstimulated (open histogram).  The cells were lysed and fixed with 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10-15 minutes at 37ºC, then permeabilized usingBD Phosflow™ Perm Buffer II (Cat. No. 558052) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-CREB (pS133) / ATF-1 (pS63) antibody.  For data analysis, lymphocytes were selected by scatter profile.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Phosflow™
Human (QC Testing), Mouse, Rat (Predicted)
Mouse BALB/c IgG1, κ
Phosphorylated Human CREB Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647122
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

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® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  4. 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.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. 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.
  7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
558434 Rev. 3
Antibody Details
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J151-21

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.

558434 Rev. 3
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
558434 Rev.3
Citations & References
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Development References (1)

  1. Shaywitz AJ, Greenberg ME. CREB: a stimulus-induced transcription factor activated by a diverse array of extracellular signals. Annu Rev Biochem. 1999; 68:821-861. (Biology).
558434 Rev. 3

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.