Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-CREB (Ser133) Blocking Peptide #1090

Description

This peptide is used to block Phospho-CREB (Ser133) Antibody #9191 reactivity.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-CREB (Ser133) Antibody #9191 by immunohistochemistry and Western Blotting.

Western Blotting

Western Blotting

Western blot analysis of extracts from forskolin and IBMX treated SK-N-MC cells, using Phospho-CREB (Ser133) Antibody #9191 with and without preincubation with phospho-peptide.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma using Phospho-CREB (Ser133) Antibody #9191 preincubated with irrelevant control peptide (left) and Phospho-CREB (Ser133) Antibody Blocking Peptide (right).

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry and Western blot protocols.

Directions for Use

For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 1 hour before allowing to react with the blot.

For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100ul total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide.

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, 371-385.
  2. Lee, M.M. et al. (1999) J. Neurosci. Res. 55, 702-712.
  3. Redmond, L. et al. (2002) Neuron 34, 999-1010.
  4. Dash, P.K. et al. (1990) Nature 345, 718-721.
  5. Yin, J.C. et al. (1994) Cell 79, 49-58.
  6. Guzowski, J.F. and McGaugh, J.L. (1997) Proc. Nat. Acad. Sci. USA 94, 2693-2698.
  7. Xing, J. et al. (1998) Mol. Cell. Biol. 18, 1946-1955.
  8. Ribar, T.J. et al. (2000) J. Neurosci. 20, RC107.
  9. Tan, Y. et al. (1996) EMBO J. 15, 4629-4642.

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