Revision 3

#7385

Store at +4C

PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H M R

UniProt ID:
#P16220

Entrez-Gene Id:
#1385

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color Storage Temp
CREB Rabbit mAb Coated Microwells5797096 tests+4C
Phospho-CREB (Ser133) Mouse Detection mAb139561 eaGreen (Lyophilized)+4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)133041 eaRed (Lyophilized)+4C
Detection Antibody Diluent1333911 mlGreen+4C
HRP Diluent1351511 mlRed+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Wash Buffer (20X)980125 ml+4C
ELISA Sample Diluent1108325 mlBlue+4C
PathScan® Sandwich ELISA Lysis Buffer (1X)701830 ml-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

Description

The PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of CREB when phosphorylated at Ser133. A CREB rabbit antibody has been coated onto the microwells. After incubation with cell lysates, CREB protein (phosphorylated and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-CREB (Ser133) mouse monoclonal detection antibody is added to detect the captured phospho-CREB (Ser133) protein. HRP-linked anti-mouse IgG is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of CREB phosphorylated at Ser133.

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

Specificity/Sensitivity

PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit #7385 detects endogenous levels of CREB protein when phosphorylated at Ser133 as shown in Figure 1. Kit sensitivity is shown in Figure 2. 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).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

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.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 3

Cell Signaling Technology Logo
Figure 1. Treatment of SK-N-MC cells with forskolin stimulates phosphorylation of CREB at Ser133, detected by the PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit #7385, but does not affect the levels of total CREB detected by PathScan® Total CREB Sandwich ELISA Kit #7390. SK-N-MC cells (80-90% confluent) were treated with 120 μM forskolin with 0.5 mM IBMX for 30 minutes and lysed. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using CREB (48H2) Rabbit mAb #9197 (left panel) or Phospho-CREB (Ser133) (1B6) Mouse mAb #9196 (right panel) are shown in the bottom figure.
ELISA Image 1: PathScan<sup>®</sup> Phospho-CREB (Ser133) Sandwich ELISA Kit
Figure 2. The relationship between the protein concentration of lysates from untreated and forskolin-treated SK-N-MC cells and the absorbance at 450 nm using the PathScan® Phospho-CREB (Ser133) Sandwich ELISA Kit is shown.
ELISA Image 2: PathScan<sup>®</sup> Phospho-CREB (Ser133) Sandwich ELISA Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.