PE Mouse Anti-CREB (pS133) / ATF-1 (pS63)

BD Phosflow™ PE Mouse Anti-CREB (pS133) / ATF-1 (pS63)

Name: PE Mouse Anti-CREB (pS133) / ATF-1 (pS63)
Brand: BD Phosflow™
SKU: 558436

Clone J151-21

(RUO)

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 (BD Phosflow™ Perm Buffer II, Cat. No. 558052) on ice for 30 minutes, and then stained with the PE 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.

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Product Details

Brand: BD Phosflow™

Reactivity: Human (QC Testing), Mouse,Rat (Predicted)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Phosphorylated Human CREB Peptide

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 20 µl

RRID: AB_647204

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: 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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

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).
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.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Lyse/Fix Buffer 5X RUO

Size 250 mL Cat No. 558049

Perm Buffer II RUO

Size 125 mL Cat No. 558052

Perm/Wash Buffer I RUO

Size 125 mL Cat No. 557885

Perm Buffer III RUO

Size 125 mL Cat No. 558050

Fixation Buffer RUO

Size 100 mL Cat No. 554655

558436 Rev. 3

Antibody Details

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.

558436 Rev. 3

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Source: Yellow-Green 488 nm, 532 nm, 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

558436 Rev.3

Citations & References

View product citations for antibody "558436" on CiteAb

Development References (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).

558436 Rev. 3

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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.