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03-199 Sigma-AldrichRIPAb+™ Phospho-eIF4E (Ser209) - RIP Validated Antibody and Primer Set, rabbit monoclonal

This RIPAb+ Phospho-eIF4E (Ser209) -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Ficha de datos de seguridad (MSDS o SDS), certificado de análisis y de calidad (CoA y CoQ), expedientes, folletos y otros documentos disponibles.

Key Applications: RNA Binding Protein Immunoprecipitation (RIP), Western Blotting Research Category: Signaling, Protein Trafficking

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Species Reactivity	Key Applications
H, M, R	RIP, WB

Description
Catalogue Number	03-199
Trade Name	RIPAb+ Upstate
Description	RIPAb+™ Phospho-eIF4E (Ser209) - RIP Validated Antibody and Primer Set, rabbit monoclonal
Overview	RIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes a negative control antibody to ensure specificity of the RIP reaction and is verified for the co-immunoprecipitation of RNA associated specifically with the immunoprecipitated RNA binding protein of interest. Where appropriate, the RIPAb+ set also includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully co-precipitating the specific RNA targets, such as messenger RNAs. The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. If a target specific assay is not provided, the RIPAb+ kit is validated using an automated microfluidics-based assay by enrichment of detectable RNA over control immunoprecipitation. eIF4E is a eukaryotic translation initiation factor that is involved in the direction of ribosomes to the cap structure of mRNA's. There are two forms of eIF4E: a free form (25 kDa) and as part of a multi-protein complex (eIF-4F). The free form seems to not be as plentiful, compared to the other initiation factors and acts as a rate limiting step of initiation.
Alternate Names	eIF-4F 25 kDa subunit eukaryotic translation initiation factor 4E eukaryotic translation initiation factor 4E-like 1 mRNA cap-binding protein
Background Information	eIF4E is a eukaryotic translation initiation factor that is involved in the direction of ribosomes to the cap structure of mRNA's. There are two forms of eIF4E: a free form (25 kDa) and as part of a multi-protein complex (eIF-4F). The free form seems to not be as plentiful, compared to the other initiation factors and acts as a rate limiting step of initiation.

References

Product Information
Format	Unpurified
Control	Includes negative control normal rabbit IgG antibody and control primers specific for the cDNA of human ACTB.
Presentation	Anti-Phospho-eIF4E (Ser209) (Rabbit Monoclonal). One vial containing 50 μL of Rabbit Monoclonal Antibody in buffer containing 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl containing 40% Glycerol, 0.01% sodium azide and 0.05% BSA. Store at -20°C. Normal Rabbit IgG. One vial containing 125 µg of Rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C. RIP Primers, ACTB, Part # CS203211. One vial containing 75 μL of 5 μM of each primer specific for human Beta-actin (ACTB) mRNA. Store at -20°C. FOR: TTG TTA CAG GAA GTC CCT TGC C REV: ATG CTA TCA CCT CCC CTG TGT G

Applications
Application	This RIPAb+ Phospho-eIF4E (Ser209) -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications	RNA Binding Protein Immunoprecipitation (RIP) Western Blotting
Application Notes	Immunoprecipitation from RIP lysate: Representative lot data. RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using 5 µg of either a normal rabbit IgG, (Cat. # PP64B), or 5 µg of Anti-phospho-eIF4E (Ser209) antibody (Cat. # CS207318). Twenty percent of the precipitated proteins (lane 1: normal mouse IgG, lane 2: phospho-eIF4E (Ser209) and HeLa whole cell lysate were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-phospho-eIF4E (Ser209) (Cat. # CS207318, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231). Arrow indicates phospho-eIF4E (Ser209). (Figure 2). Western Blot Analysis: Representative lot data. 293 cell lysate was resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-phospho-eIF4E (Ser209), clone EP2151Y (1:10,000 dilution); untreated (A) and treated with AP (B). Proteins were visualized using a Goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-eIF4E (Ser209) (~25 kDa). (Figure 3).

Biological Information
Immunogen	Synthetic peptide corresponding to residues surrounding phosphorylated Ser209 of human eIF4E.
Epitope	Phosphorylated Ser209
Host	Rabbit
Specificity	This antibody recognizes eIF4E phosphorylated at Ser209.
Isotype	IgG
Species Reactivity	Human Mouse Rat
Species Reactivity Note	Human, mouse, and rat.
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_001959.1
Entrez Gene Summary	All eukaryotic cellular mRNAs are blocked at their 5-prime ends with the 7-methylguanosine cap structure, m7GpppX (where X is any nucleotide). This structure is involved in several cellular processes including enhanced translational efficiency, splicing, mRNA stability, and RNA nuclear export. EIF4E is a eukaryotic translation initiation factor involved in directing ribosomes to the cap structure of mRNAs. It is a 24-kD polypeptide that exists as both a free form and as part of a multiprotein complex termed EIF4F. The EIF4E polypeptide is the rate-limiting component of the eukaryotic translation apparatus and is involved in the mRNA-ribosome binding step of eukaryotic protein synthesis. The other subunits of EIF4F are a 50-kD polypeptide, termed EIF4A (see MIM 601102), that possesses ATPase and RNA helicase activities, and a 220-kD polypeptide, EIF4G (MIM 600495) (Rychlik et al., 1987 [PubMed 3469651]).
Gene Symbol	CBP EIF4E1 EIF4EL1 EIF4F MGC111573 eIF-4E eIF4E
Modifications	Phosphorylation
Purification Method	Unpurified
UniProt Number	P06730
UniProt Summary	FUNCTION: Recognizes and binds the 7-methylguanosine-containing mRNA cap during an early step in the initiation of protein synthesis and facilitates ribosome binding by inducing the unwinding of the mRNAs secondary structures. SUBUNIT STRUCTURE: eIF4F is a multi-subunit complex, the composition of which varies with external and internal environmental conditions. It is composed of at least EIF4A, EIF4E and EIF4G1/EIF4G3. EIF4E is also known to interact with other partners. The interaction with EIF4ENIF1 mediates the import into the nucleus. Nonphosphorylated EIF4EBP1, EIF4EBP2 and EIF4EBP3 compete with EIF4G1/EIF4G3 to interact with EIF4E; insulin stimulated MAP-kinase (MAPK1 and MAPK3) phosphorylation of EIF4EBP1 causes dissociation of the complex allowing EIF4G1/EIF4G3 to bind and consequent initiation of translation. Rapamycin can attenuate insulin stimulation, mediated by FKBPs. Interacts mutually exclusive with EIF4A1 and EIF4A2. Interacts with NGDN and PIWIL2 By similarity. Interacts with Lassa virus Z protein. PTM: Phosphorylation increases the ability of the protein to bind to mRNA caps and to form the eIF4F complex. SEQUENCE SIMILARITIES: Belongs to the eukaryotic initiation factor 4E family. CAUTION: Was originally thought to be phosphorylated on Ser-53 ; this was later shown to be wrong
Molecular Weight	~25 kDa

Physicochemical Information

Dimensions

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Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	RNA Binding Protein Immunoprecipitation: Representative lot data. RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal rabbit IgG or 5 µg of Anti-phospho-eIF4E (Ser209) antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of phospho-eIF4E (Ser209)-associated RNA was verified by qPCR using RIP Primers, ACTB, (Figure 1). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.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

RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal rabbit IgG or 5 µg of Anti-phospho-eIF4E (Ser209) antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of phospho-eIF4E (Ser209)-associated RNA was verified by qPCR using RIP Primers, ACTB, (Figure 1).
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.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 -20ºC from date of receipt. Handling Recommendations: Upon receipt 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.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at -20ºC from date of receipt.
Handling Recommendations: Upon receipt 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 Size	10 assays
Material Package	10 assays per set. Recommended use: ~5 μL of antibody per RIP (dependent upon biological context).

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Número de referencia	GTIN
03-199	04053252558245

Documentation

RIPAb+™ Phospho-eIF4E (Ser209) - RIP Validated Antibody and Primer Set, rabbit monoclonal Ficha datos de seguridad (MSDS)

Título
Ficha técnica de seguridad del material (MSDS)

RIPAb+™ Phospho-eIF4E (Ser209) - RIP Validated Antibody and Primer Set, rabbit monoclonal Certificados de análisis

Cargo	Número de lote
RIPAb+ Phospho-eIF4E (Ser209) - 2161196	2161196
RIPAb+ Phospho-eIF4E (Ser209) - NRG1889585	NRG1889585
RIPAb+ Phospho-eIF4E (Ser209)	2467155
RIPAb+ Phospho-eIF4E (Ser209) -2489248	2489248

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Número de referencia	Descripción
07-823	Anti-phospho eIF4E (Ser209) Antibody

Productos relacionados por: Brand Facete

Categorías

Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies

Life Science Research > Antibodies and Assays > Primary Antibodies

Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies

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