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17-10061 ChIPAb+ E2F-1 - ChIP Validated Antibody and Primer Set

This ChIPAb+ E2F-1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Applications: Immunoprecipitation, ChIP-seq, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Transcription Factors
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17-10061
25 assays  25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
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      Overview

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      100% Performance Guaranteed

      Key Spec Table

      Species ReactivityKey Applications
      H, XnIP, ChIP-seq, ChIP
      Description
      Catalogue Number17-10061
      Brand Family Upstate
      Trade Name
      • ChIPAb+
      • Upstate
      DescriptionChIPAb+ E2F-1 - 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+ E2F-1 set includes the E2F-1 antibody, a negative control normal mouse IgG, and qPCR primers which amplify a 100 bp region of human CDC2 promoter. The E2F-1 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of E2F-1-associated chromatin.
      Alternate Names
      • Transcription factor E2F1
      • Retinoblastoma-binding protein 3
      • PRB-binding protein E2F-1
      Background InformationE2F-1 was identified as a cellular protein with DNA binding activity associated with the adenovirus E2 gene promoter. The original member of the E2F family of transcription factors, now consisting of five, was identified through its association with the retinoblastoma protein, which controls its activity. E2F-1 is cell cycle regulated with very low levels in early G1 then increasing levels as cells move from G1 to S with highest levels of protein at the G1/S boundary, which is consistent with its role in S-phase entry.
      Product Information
      FormatPurified
      Control
      • Includes negative control normal mouse IgG and primers specific for human CDC2 promoter.
      PresentationAnti-E2F-1 (mouse monoclonal). One vial containing 25 µg of protein G purified monoclonal mixed IgG in 25 µL of buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl with 0.05% sodium azide. Frozen solution. Store at -20°C.

      Nornal Mouse IgG. One vial containing 25 µg of purified IgG in 25 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.

      ChIP Primers, CDC2 promoter. One vial containing 75 μL of 5 μM each primer specific for human CDC2 promoter. Store at -20°C.
      FOR: CGC CCT TTC CTC TTT CTT TC
      REV: ATC GGG TAG CCC GTA GAC TT
      Quality LevelMQ100
      Applications
      ApplicationThis ChIPAb+ E2F-1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • Immunoprecipitation
      • ChIP-seq
      • Chromatin Immunoprecipitation (ChIP)
      Application NotesChromatin Immunoprecipitation:
      Representative lot data.
      Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Mouse IgG or 1 µL of Anti-E2F-1 and the Magna ChIP™ G Kit (Cat. # 17-611).
      Successful immunoprecipitation of E2F-1-associated DNA fragments was verified by qPCR using ChIP Primers, human CDC2 promoter as a positive locus, and an intergenic region near human CALML5 as a negative locus. (Please see figures). 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™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

      Western Blot Analysis:
      Representative lot data.
      HeLa nuclear extract was resolved by electrophoresis, transferred to PVDF and probed with Anti-E2F-1 (1.0 µg/mL).
      Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection
      system.
      Arrow indicates E2F-1 (~55 kDa) (Please see figures).


      Immunoprecipitation: 4 μg of a previous lot of this antibody immunoprecipitated E2F-1 from HeLa nuclear extract.

      Electrophoretic Mobility Shift Assay (EMSA): A previous lot of the antibody has been shown to gel shift the E2F-1/DP-1/DNA complex.
      Biological Information
      ImmunogenGST fusion protein corresponding to full-length human E2F-1.
      Epitopea.a. 1-89 & 342-386
      HostMouse
      SpecificityThe epitopes have been mapped to amino acids 1-89 and to amino acids 342-386.
      IsotypeIgG
      Species Reactivity
      • Human
      • Xenopus
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThe protein encoded by this gene is a member of the E2F family of transcription factors. The E2F family plays a crucial role in the control of cell cycle and action of tumor suppressor proteins and is also a target of the transforming proteins of small DNA tumor viruses. The E2F proteins contain several evolutionally conserved domains found in most members of the family. These domains include a DNA binding domain, a dimerization domain which determines interaction with the differentiation regulated transcription factor proteins (DP), a transactivation domain enriched in acidic amino acids, and a tumor suppressor protein association domain which is embedded within the transactivation domain. This protein and another 2 members, E2F2 and E2F3, have an additional cyclin binding domain. This protein binds preferentially to retinoblastoma protein pRB in a cell-cycle dependent manner. It can mediate both cell proliferation and p53-dependent/independent apoptosis.
      Gene Symbol
      • E2F1
      • RBAP-1
      • E2F-1
      • RBP3
      • RBBP3
      • RBBP-3
      • PBR3
      Purification MethodProtein G Purified
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q01094 # Transcription activator that binds DNA cooperatively with dp proteins through the E2 recognition site, 5'-TTTC[CG]CGC- 3' found in the promoter region of a number of genes whose products are involved in cell cycle regulation or in DNA replication. The DRTF1/E2F complex functions in the control of cell-cycle progression from G1 to S phase. E2F-1 binds preferentially RB1 protein, in a cell-cycle dependent manner. It can mediate both cell proliferation and p53-dependent apoptosis.
      SIZE: 437 amino acids; 46920 Da
      SUBUNIT: Component of the DRTF1/E2F transcription factor complex. Forms heterodimers with DP family members. The E2F-1 complex binds specifically hypophosphorylated retinoblastoma protein RB1. During the cell cycle, RB1 becomes phosphorylated in mid-to-late G1 phase, detaches from the DRTF1/E2F complex, rendering E2F transcriptionally active. Viral oncoproteins, notably E1A, T- antigen and HPV E7, are capable of sequestering RB protein, thus releasing the active complex. Interacts with TRRAP, which probably mediates its interaction with histone acetyltransferase complexes, leading to transcription activation. Binds TOPBP1 and EAPP. Interacts with ARID3A.
      SUBCELLULAR LOCATION: Nucleus.
      PTM: Phosphorylated by CDK2 and cyclin A-CDK2 in the S-phase.
      SIMILARITY: SwissProt: Q01094 ## Belongs to the E2F/DP family.
      Molecular Weight~55 kDa
      Product Usage Statements
      Quality AssuranceChromatin Immunoprecipitation:
      Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Mouse IgG or 1 µL of Anti-E2F-1 and the Magna ChIP™ G Kit (Cat. # 17-611).
      Successful immunoprecipitation of E2F-1 associated DNA fragments was verified by qPCR using ChIP Primers, human CDC2 promoter (Please see figures).

      Please refer to the EZ-Magna ChIP™ G (Cat. # 17-409) 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.
      Packaging Information
      Material Size25 assays
      Material Package25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
      Global Trade Item Number
      Catalogue Number GTIN
      17-10061 04053252412899