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17-10061 Sigma-AldrichChIPAb+ 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.

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Key Applications: Immunoprecipitation, ChIP-seq, Chromatin Immunoprecipitation (ChIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: Transcription Factors

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Key Spec Table

Species Reactivity	Key Applications
H, Xn	IP, ChIP-seq, ChIP

Description
Catalogue Number	17-10061
Brand Family	Upstate
Trade Name	ChIPAb+ Upstate
Description	ChIPAb+ E2F-1 - ChIP Validated Antibody and Primer Set
Overview	All 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 Information	E2F-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.

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Product Information
Format	Purified
Control	Includes negative control normal mouse IgG and primers specific for human CDC2 promoter.
Presentation	Anti-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 Level	MQ100

Applications
Application	This 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 Notes	Chromatin 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
Immunogen	GST fusion protein corresponding to full-length human E2F-1.
Epitope	a.a. 1-89 & 342-386
Host	Mouse
Specificity	The epitopes have been mapped to amino acids 1-89 and to amino acids 342-386.
Isotype	IgG
Species Reactivity	Human Xenopus
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_005216
Entrez Gene Summary	The 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 Method	Protein G Purified
UniProt Number	Q01094
UniProt Summary	FUNCTION: 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

Physicochemical Information

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Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	Chromatin 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.

Product Usage Statements

Quality Assurance

Chromatin 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 Conditions	Stable 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 Size	25 assays
Material Package	25 assays per set. Recommended use: ~1 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).

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Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
17-10061	04053252412899

Documentation

ChIPAb+ E2F-1 - ChIP Validated Antibody and Primer Set MSDS

Title
Safety Data Sheet (SDS)

ChIPAb+ E2F-1 - ChIP Validated Antibody and Primer Set Certificates of Analysis

Title	Lot Number
ChIPAb+ E2F-1 -2491691	2491691
ChIPAb+ E2F-1 - 2110515	2110515
ChIPAb+ E2F-1 - 2490685	2490685
ChIPAb+ E2F-1 - 3254861	3254861
ChIPAb+ E2F-1 - 3495701	3495701
ChIPAb+ E2F-1 - 3495710	3495710
ChIPAb+ E2F-1 - 3521158	3521158
ChIPAb+ E2F-1 - 3533144	3533144
ChIPAb+ E2F-1 - 3698766	3698766
ChIPAb+ E2F-1 - 3870091	3870091

References

Reference overview	Pub Med ID
miR-148a is Associated with Obesity and Modulates Adipocyte Differentiation of Mesenchymal Stem Cells through Wnt Signaling. Shi, C; Zhang, M; Tong, M; Yang, L; Pang, L; Chen, L; Xu, G; Chi, X; Hong, Q; Ni, Y; Ji, C; Guo, X Scientific reports 5 9930 2015 Show Abstract Obesity results from numerous, interacting genetic, behavioral, and physiological factors. Adipogenesis is partially regulated by several adipocyte-selective microRNAs (miRNAs) and transcription factors that regulate proliferation and differentiation of human adipose-derived mesenchymal stem cells (hMSCs-Ad). In this study, we examined the roles of adipocyte-selective miRNAs in the differentiation of hMSCs-Ad to adipocytes. Results showed that the levels of miR-148a, miR-26b, miR-30, and miR-199a increased in differentiating hMSCs-Ad. Among these miRNAs, miR-148a exhibited significant effects on increasing PPRE luciferase activity (it represents PPAR-dependent transcription, a major factor in adipogenesis) than others. Furthermore, miR-148a expression levels increased in adipose tissues from obese people and mice fed high-fat diet. miR-148a acted by suppressing its target gene, Wnt1, an endogenous inhibitor of adipogenesis. Ectopic expression of miR-148a accelerated differentiation and partially rescued Wnt1-mediated inhibition of adipogenesis. Knockdown of miR-148a also inhibited adipogenesis. Analysis of the upstream region of miR-148a locus identified a 3 kb region containing a functional cAMP-response element-binding protein (CREB) required for miR-148a expression in hMSCs-Ad. The results suggest that miR-148a is a biomarker of obesity in human subjects and mouse model, which represents a CREB-modulated miRNA that acts to repress Wnt1, thereby promoting adipocyte differentiation.	26001136

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