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03-249 Sigma-AldrichRIPAb+™ Ago1 Antibody

This RIPAb+ Ago1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

RIPAb+™ Ago1 Antibody MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

SDS
CoA
Brochures

Key Applications: Western Blotting, RNA Binding Protein Immunoprecipitation (RIP) Research Category: Epigenetics & Nuclear Function Research Sub-Category: RNA Binding Protein (RBP)

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Key Spec Table

Species Reactivity	Key Applications
H	WB, RIP

Description
Catalogue Number	03-249
Trade Name	RIPAb+ Upstate
Description	RIPAb+™ Ago1 Antibody
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. Ago1 (Eukaryotic translation initiation factor 2C 1) is a member of the argonaute family. It contains a PAZ and a PIWI domain, and is required for G1 arrest and mating in response to nitrogen starvation. It may also contribute to cell cycle control by regulating cytokinesis. Ago1 is also needed for the production of mature miRNA that affects miRNA-directed RNA cleavage and is therefore important for heterchromatin formation and accurate chromosome segregation.
Alternate Names	hAgo1 Argonaute1 eIF-2C 1 eIF2C 1 Q99 GERP95 DKFZp686M13167
Background Information	Ago1 (Eukaryotic translation initiation factor 2C 1) belongs to the argonaute family and contains 1 PAZ and 1 Piwi domain. It is required for G1 arrest and mating in response to nitrogen starvation. It is also known as a cell cycle control protein through its regulation of cytokinesis and its related cell cycle checkpoints, which occur downstream of dcr1. It is required, indirectly, for regulated hyperphosphorylation of cdc2. Ago 1 also has a role in the RNA interference (RNAi) pathway, which is important for heterochromatin formation and accurate chromosome segregation. It has been shown to be non-essential for siRNA-directed target RNA cleavage, but is needed for the production of mature miRNA that affects miRNA-directed RNA cleavage.

References

Product Information
Format	Purified
Control	Includes normal mouse IgG and primers specific for human IGF2 mRNA.
Presentation	Anti-Ago1 (Mouse Monoclonal). One vial containing 50 µg protein G purified monoclonal in buffer containing 0.1 M Tris-glycine (pH 7.4), 150mM NaCl, 0.05% sodium azide before the addition of glycerol to 30%. Store at -20° C. Concentration: 0.7 mg/mL Normal mouse IgG. One vial containing 125 µg purified mouse IgG in 125 µL storage buffer containing 0.1% sodium azide. Store at -20°C. RIP Primer IGF2. One vial containing 75 μL of 5 μM of each primer specific for human IGF2 mRNA. Store at -20°C. FOR: GCG GCT TCT ACT TCA GCA G REV: CAG GTG TCA TAT TGG AAG AAC
Quality Level	MQ100

Applications
Application	This RIPAb+ Ago1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications	Western Blotting RNA Binding Protein Immunoprecipitation (RIP)
Application Notes	RNA Binding Protein Immunoprecipitation (microRNA): Representative lot data. RIP Lysate prepared from 293 cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG (Cat. # CS200621), or Anti-Ago1 antibody (Cat. # CS207365) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of Ago 1-associated microRNA was verified by TaqMan® microRNA assay, hsa-miR-15a. (Figure2). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details. Western Blot Analysis: Representative lot data. HeLa cell lysate was probed with Anti-Ago 1, clone 4G7-E12 (0.1 µg/mL). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Ago 1 (~97 kDa).

Biological Information
Immunogen	GST-tagged recombinant protein corresponding to human Ago1
Clone	4G7-E12
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Mouse
Specificity	This antibody has been shown by an independent laboratory to not cross react with Ago2, Ago3, or Ago4 (Azuma-Mukai A et al. (2008) Proc. Natl. Acad. Sci. USA 105: 7964-7969).
Isotype	IgG2aκ
Species Reactivity	Human
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NP_036331
Entrez Gene Summary	This gene encodes a member of the Argonaute family of proteins which play a role in RNA interference. The encoded protein is highly basic, and contains a PAZ domain and a PIWI domain. It may interact with dicer1 and play a role in short-interfering-RNA-mediated gene silencing. This gene is located on chromosome 1 in a cluster of closely related family members including argonaute 3, and argonaute 4.
Gene Symbol	EIF2C1 AGO1
Purification Method	Protein G Purified
UniProt Number	Q9UL18
UniProt Summary	FUNCTION: Required for RNA-mediated gene silencing (RNAi). Binds to short RNAs such as microRNAs (miRNAs) or short interfering RNAs (siRNAs), and represses the translation of mRNAs which are complementary to them. Lacks endonuclease activity and does not appear to cleave target mRNAs. Also required for transcriptional gene silencing (TGS) of promoter regions which are complementary to bound short antigene RNAs (agRNAs). SUBUNIT STRUCTURE: Interacts with DDB1, DDX6, DDX20, DDX47, DHX9, DICER1, EIF2C2/AGO2, GEMIN4, ILF3, IMP8, MOV10, PRMT5, SART3, TARBP2, TNRC6B and UPF1. SUBCELLULAR LOCATION: Cytoplasm > P-body.
Molecular Weight	~97 kDa

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	RNA Binding Protein Immunoprecipitation: Representative lot data RIP Lysate prepared from 293 cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG, or 5 µg Anti-Ago1 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of Argonaute 1-associated RNA was verified by qPCR using RIP Primers IGF2, (Figure 1). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) 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 293 cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG, or 5 µg Anti-Ago1 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Argonaute 1-associated RNA was verified by qPCR using RIP Primers IGF2, (Figure 1).
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) 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. 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 -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 Size	10 assays
Material Package	10 assays per set. Recommended use: ~5 µg of antibody per RIP (dependent upon biological context).

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
03-249	04053252439414

Documentation

RIPAb+™ Ago1 Antibody SDS

Title
Safety Data Sheet (SDS)

RIPAb+™ Ago1 Antibody Certificates of Analysis

Title	Lot Number
RIPAb+ Ago1 - 2262704	2262704
RIPAb+ Ago1 - Q1999623	Q1999623
RIPAb+ Ago1 - 3304135	3304135
RIPAb+ Ago1 - 3431390	3431390
RIPAb+ Ago1 - 3691558	3691558
RIPAb+ Ago1 - 3878890	3878890
RIPAb+ Ago1 - 4087652	4087652
RIPAb+ Ago1 -2797334	2797334
RIPAb+ Ago1 Monoclonal Antibody/Primer Set	3085863
RIPAb+ Ago1 Monoclonal Antibody/Primer Set	3036800

Brochure

Title
New Products: Volume 3, 2012

Catalogue Number	Description
04-083	Anti-Ago1 Antibody, clone 6D8.2
MABE1121	Anti-Ago1 Antibody, clone 4B8
MABE143	Anti-Ago1 Antibody, clone 4G7-E12

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03-249 Sigma-AldrichRIPAb+™ Ago1 Antibody

This RIPAb+ Ago1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Recommended Products

Overview

Special Offers

Key Spec Table

Documentation

RIPAb+™ Ago1 Antibody SDS

RIPAb+™ Ago1 Antibody Certificates of Analysis

Brochure

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