Revision 7

#42054Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H M R Mk

UniProt ID:

#P16220

Entrez-Gene Id:

#1385

Cell Signaling Technology

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3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color
FastScan ELISA Microwell Strip Plate, 96 Well 53257 96 tests
Phospho-CREB (Ser133) Mouse Capture mAb 59667 1 ea Green (Lyophilized)
CREB Rabbit HRP-linked mAb 76974 1 ea Red (Lyophilized)
FastScan ELISA Capture Antibody Diluent 16076 3 ml Green
FastScan ELISA HRP Antibody Diluent 28120 3 ml
TMB Substrate 7004 11 ml
STOP Solution 7002 11 ml
Sealing Tape 54503 1 ea
ELISA Wash Buffer (20X) 9801 25 ml
FastScan ELISA Cell Extraction Buffer (5X) 69905 10 ml
FastScan ELISA Cell Extraction Enhancer Solution (50X) 25243 1 ml
FastScan ELISA Kit #42054 Positive Control Type 1 97186 1 ea

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The FastScan Phospho-CREB (Ser133) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of CREB when phosphorylated at Ser133. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with phospho-CREB (Ser133) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of phospho-CREB (Ser133).

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

The FastScan Phospho-CREB (Ser133) ELISA Kit detects endogenous levels of CREB when phosphorylated at Ser133 as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.This kit also detects the phosphorylated form of the CREB-related protein ATF-1.

Background

CREB is a bZIP transcription factor that activates target genes through cAMP response elements. CREB is able to mediate signals from numerous physiological stimuli, resulting in regulation of a broad array of cellular responses. While CREB is expressed in numerous tissues, it plays a large regulatory role in the nervous system. CREB is believed to play a key role in promoting neuronal survival, precursor proliferation, neurite outgrowth, and neuronal differentiation in certain neuronal populations (1-3). Additionally, CREB signaling is involved in learning and memory in several organisms (4-6). CREB is able to selectively activate numerous downstream genes through interactions with different dimerization partners. CREB is activated by phosphorylation at Ser133 by various signaling pathways, including Erk, Ca2+, and stress signaling. Some of the kinases involved in phosphorylating CREB at Ser133 are p90RSK, MSK, CaMKIV, and MAPKAPK-2 (7-9).

  1. Lonze, B.E. et al. (2002) Neuron 34, 371-85.
  2. Lee, M.M. et al. (1999) J Neurosci Res 55, 702-12.
  3. Redmond, L. et al. (2002) Neuron 34, 999-1010.
  4. Dash, P.K. et al. (1990) Nature 345, 718-21.
  5. Yin, J.C. et al. (1994) Cell 79, 49-58.
  6. Guzowski, J.F. and McGaugh, J.L. (1997) Proc Natl Acad Sci USA 94, 2693-8.
  7. Xing, J. et al. (1998) Mol Cell Biol 18, 1946-55.
  8. Ribar, T.J. et al. (2000) J Neurosci 20, RC107.
  9. Tan, Y. et al. (1996) EMBO J 15, 4629-42.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 7
    #42054

    FastScan Phospho-CREB (Ser133) ELISA Kit

    FastScan™ Phospho-CREB (Ser133) ELISA Kit: Image 1 Expand Image
    Figure 1. Treatment of NIH/3T3 cells with PDGF stimulates phosphorylation of CREB at Ser133 but does not affect the level of total CREB. The relationship between lysate protein concentration from untreated and PDGF-treated NIH/3T3 cells and the absorbance at 450 nm using the FastScan Phospho-CREB (Ser133) ELISA Kit #42054 is shown in the upper figure. The corresponding western blots using CREB antibody (left panel) and phospho-CREB (Ser133) antibody (right panel) are shown in the lower figure. After serum starvation, NIH/3T3 cells were treated with 100 ng/ml PDGF #8912 for 5 minutes at 37°C and then lysed.