Human/Mouse/Rat Phospho-CREB (S133) Cell-Based ELISA

Discontinued Product

KCB2510 has been discontinued.
View all CREB products.
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Human/Mouse/Rat Phospho-CREB (S133) Cell-Based ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Alternatively
* Alternatively, this assay can be run over two days, which requires 2 hours 30 mins (day 1) and 4 hours (day 2).

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

* Alternatively, this assay can be run over two days, which requires 2 hours 30 mins (day 1) and 4 hours (day 2).


PRODUCT SUMMARY

R&D Systems Cell-Based ELISAs simultaneously measure the levels of two proteins in the same microplate well in whole, fixed cells, eliminating the need for lysate preparation. These simple, sensitive assays can be used with both adherent and non-adherent cells to analyze protein phosphorylation or assess total protein levels.

Cell-Based ELISA Kits are available in two formats: phospho-protein kits contain antibodies to measure both the phosphorylated and the total protein, while total protein kits contain antibodies to the protein of interest and a second housekeeping protein. These two proteins are simultaneously detected in the same microplate well, allowing signals derived from the target protein to be normalized to that of the second protein. This permits corrections for well-to-well variation and allows target protein levels to be accurately assessed and compared across multiple samples. This Phospho- Cell-Based ELISA Kit employs fluorometric detection that is compatible with standard microplate fluorometers.


PRODUCT FEATURES

  • No lysate preparation required
  • Results with as few as 10,000 cells per well
  • Measure two proteins simultaneously in the same well
  • Ideal for measuring phosphorylation
  • Design allows for normalization of well-to-well variations
  • Complete kits require minimal optimization
  • Suitable for high-throughput analysis of non-adherent cells
  • Excellent alternative to Western blot and/or sandwich ELISA


KIT CONTENTS

  • 96-well Microplate
  • Primary antibody for target protein
  • Primary antibody for normalization protein
  • HRP-conjugated secondary antibody
  • AP-conjugated secondary antibody
  • HRP Substrate F1
  • AP Substrate F2
  • Blocking Buffer
  • Fixing Solution 1
  • Wash Buffer
  • Plate Sealers


OTHER REAGENTS REQUIRED

  • 30% H2O2 (Sigma, Catalog # H1009). Refer to MSDS prior to use
  • 1X PBS (Irvine Scientific, Catalog # 9240)
  • Deionized or distilled water
  • Pipettes and pipette tips
  • Multi-channel pipette for washing
  • Cell culture incubator
  • Microfuge tubes
  • Orbital shaker
  • Fluorescence plate reader with two channels: excitation 540 nm/emission 600 nm and excitation 360 nm/emission 450 nm

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CREB

The cAMP Response Element-Binding Protein (CREB) belongs to the bZIP superfamily of transcription factors, containing a basic domain that mediates DNA binding, and a leucine zipper domain that facilitates dimerization. Within the promoter of target genes, CREB dimers bind cAMP response elements, defined by the palindromic consensus sequence TGACGTCA. When phosphorylated at S133, CREB also binds the coactivator CREB binding protein (CBP), which enhances transcription by acetylating histones to facilitate chromatin unraveling.

Long Name:
cAMP Response Element-binding Protein
Entrez Gene IDs:
1385 (Human); 12912 (Mouse)
Alternate Names:
active transcription factor CREB; cAMP responsive element binding protein 1; cAMP-response element-binding protein-1; cAMP-responsive element-binding protein 1; CREB; CREB1; CREB-1; cyclic AMP-responsive element-binding protein 1; MGC9284; transactivator protein

FAQs

No product specific FAQs exist for this product, however you may

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