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17-10131 ChIPAb+™ Phospho-CREB (Ser133) - ChIP Validated Antibody and Primer Set

17-10131
25 assays  25 assays per set. Recommended use: ~4 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, R, M, HtEMSA, IHC, IP, WB, ChIP
      Description
      Catalogue Number17-10131
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+™ Phospho-CREB (Ser133) - ChIP Validated Antibody and Primer Set
      OverviewAll ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
      The ChIPAb+ Phospho-CREB (Ser133) set includes the Phospho-CREB (Ser133) antibody, a negative control normal rabbit IgG, and qPCR primers which amplify a 64 bp region of human cFos CRE. The Phospho-CREB (Ser133) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Phospho-CREB (Ser133)-associated chromatin.
      Alternate Names
      • active transcription factor CREB
      • cAMP responsive element binding protein 1
      • cAMP-response element-binding protein-1
      • transactivator protein
      Background InformationCREB is a β-ZIP 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. CREB promotes outgrowth and differentiation as a mediator of neurotrophin pathways. 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.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control normal rabbit IgG and primers specific for human cFOS CRE.
      PresentationAnti-phospho-CREB (Ser133) (rabbit polyclonal), Part No. CS204400. One vial containing 100 µg of affinity purified rabbit serum in 0.1 M Tris-Glycine (pH 7.4) 0.15 M NaCl, and 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C.
      Normal Rabbit IgG, Part No. PP64B. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide.
      Store at -20°C.
      ChIP Primers cFos CRE, Part No. CS203203. One vial containing 75 μL of 5 μM of each primer specific for the cFos CRE. Store at -20°C.
      FOR: GGC CCA CGA GAC CTC TGA GAC A
      REV: GCC TTG GCG CGT GTC CTA ATC T
      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ Phospho-CREB (Ser133) -ChIP Validated Antibody & Primer Set (Antibody ChIP) conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Electrophoretic Mobility Shift Assay
      • Immunohistochemistry
      • Immunoprecipitation
      • Western Blotting
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from 293T cells (5 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 4 µg of Normal Rabbit IgG, (Part No. P64B), or 4 µg of Anti-Phospho-CREB (Ser133) antibody (Part No. CS204400) and the Magna ChIP™ A Kit (Cat. # 17-610).
      Successful immunoprecipitation of Phospho-CREB (Ser133) antibody associated DNA fragments was verified by qPCR using ChIP Primers cFos CRE (Part No. CS203203) as a positive locus, and cFos downstream 4Kb as a negative locus (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      Western Blot Analysis:
      Representative lot data.
      Untreated (lane 1) and Forskolin-treated (Lane 2) NIH/3T3 lysates were resolved by SDS-PAGE, transferred to PVDF, and probed with anti-phospho-CREB (Ser133) (1:1,000 dilution).
      Proteins were visualized using a donkey anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
      Arrow indicates phosphorylated CREB (Figure 3).
      Immunoprecipitation:
      10 µL from a representative lot of this antibody was shown to immunoprecipitate phosphorylated CREB from 500 µg of cells treated with 50 mM forskolin.
      Immunohistochemistry: A 1:1000 dilution of a representative lot detected phosphorylated CREB in formalin-fixed paraffin-embedded normal and ischemic rat brain sections.
      Electrophoretic Mobility Shift:
      A representative lot of this antibody has been shown to gel shift by an independent laboratory.
      Biological Information
      ImmunogenKLH-conjugated synthetic peptide corresponding to the amino acids including and encompassing phosphorylated Ser133 of CREB.
      Epitopephospho-Ser133
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostRabbit
      SpecificityRecognizes p43 phosphorylated CREB. May also recognize p30 and p38 kDa proteins. The p30 & p38 proteins may be phosphorylated CREM and ATF-1 respectively, since the two proteins share significant homology to the phosphorylated CREB immunizing peptide.
      IsotypeIgG
      Species Reactivity
      • Human
      • Rat
      • Mouse
      • Hamster
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a transcription factor that is a member of the leucine zipper family of DNA binding proteins. This protein binds as a homodimer to the cAMP-responsive element, an octameric palindrome. The protein is phosphorylated by several protein kinases, and induces transcription of genes in response to hormonal stimulation of the cAMP pathway. Alternate splicing of this gene results in two transcript variants encoding different isoforms.
      Gene Symbol
      • CREB1
      • CREB
      • MGC9284
      Modifications
      • Phosphorylation
      Purification MethodPurified by protein A-Sepharose followed by immunoadsorbant chromatography using an unphosphorylated CREB affinity gel column.
      UniProt Number
      UniProt SummaryFUNCTION:This protein binds the cAMP response element (CRE), a sequence present in many viral and cellular promoters. CREB stimulates transcription on binding to the CRE. Transciption activation is enhanced by the TORC coactivators which act independently of Ser-133 phosphorylation. Implicated in synchronization of circadian rhythmicity.
      SUBUNIT: Interacts with PPRC1. Binds DNA as a dimer. This dimer is stabilized by magnesium ions. Interacts, through the bZIP domain, with the coactivators TORC1/CRTC1, TORC2/CRTC2 and TORC3/CRTC3. When phosphorylated on Ser-133, binds CREBBP By similarity.
      SUBCELLULAR: Nucleus.
      PMT:Stimulated by phosphorylation. Phosphorylation of both Ser-133 and Ser-142 in the SCN regulates the activity of CREB and participates in circadian rhythm generation. Phosphorylation of Ser-133 allows CREBBP binding By similarity. Phosphorylated upon DNA damage, probably by ATM or ATR.
      SIMILARITY: Belongs to the bZIP family.
      Contains 1 bZIP domain.
      Contains 1 KID (kinase-inducible) domain
      Molecular WeightDetects CREB only when phosphorylated on Ser133 (~43 kDa). May recognize ATF-1 (~38 kDa) and CREM (~30 kDa) as the two proteins share significant homology to the phosphorylated immunizing peptide.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from 293T cells (5 X 10E6 cell equivalents per IP) was subjected to chromatin immunoprecipitation using either 4 μg of a Normal Rabbit IgG , or 4 μg of Anti-Phospho-CREB antibody and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of Phospho-CREB (Ser133) associated DNA fragments was verified by qPCR using ChIP Primers cFos CRE (Figure 1).
      Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~4 μg of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      ChIPAb+™ Phospho-CREB (Ser133) - ChIP Validated Antibody and Primer Set Certificates of Analysis

      TitleLot Number
      ChIPAb+ Phospho-CREB (Ser133) - 2119017 2119017
      ChIPAb+ Phospho-CREB (Ser133) - 2006391 2006391
      ChIPAb+ Phospho-CREB (Ser133) - 2035083 2035083
      ChIPAb+ Phospho-CREB (Ser133) - NRG1810821 NRG1810821
      ChIPAb+™ Phospho-CREB (Ser133) - 3006573 3006573
      ChIPAb+™ Phospho-CREB (Ser133) - 3190991 3190991
      ChIPAb+™ Phospho-CREB (Ser133) - 3433785 3433785
      ChIPAb+™ Phospho-CREB (Ser133) - 3563626 3563626
      ChIPAb+™ Phospho-CREB (Ser133) - 3828306 3828306
      ChIPAb+™ Phospho-CREB (Ser133) - 3843809 3843809

      References

      Reference overviewPub Med ID
      Molecular mechanisms mediating a deficit in recall of fear extinction in adult mice exposed to cocaine in utero.
      Kabir, ZD; Katzman, AC; Kosofsky, BE
      PloS one  8  e84165  2013

      Show Abstract
      24358339 24358339

      Brochure

      Title
      Advance your Epigenetics Research

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      Categories

      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies
      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > Chromatin Immunoprecipitation (ChIP) > ChIP Validated Antibodies