03-248
RIPAb+ pan Ago - RIP Validated Antibody and Primer Set
clone 2A8, from mouse
Synonyme(s) :
Eukaryotic translation initiation factor 2C, 2, Protein argonaute-2, Ago2, Protein slicer, Argonaute2, PAZ Piwi domain protein, Ago4, Argonaute4, Protein argonaute-4
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About This Item
Code UNSPSC :
12352203
eCl@ss :
32160702
Nomenclature NACRES :
NA.32
Produits recommandés
Source biologique
mouse
Niveau de qualité
Forme d'anticorps
purified immunoglobulin
Clone
2A8, monoclonal
Espèces réactives
human
Fabricant/nom de marque
RIPAb+
Upstate®
Technique(s)
RIP: suitable
immunocytochemistry: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
Isotype
IgG1κ
Numéro d'accès NCBI
Numéro d'accès UniProt
Conditions d'expédition
dry ice
Description générale
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.
Argonaute (Ago) proteins are a family of proteins of ~95 kDa that bind directly to mature miRNAs. In mammals, there are four Ago proteins (Ago1-4). This family of proteins is a central component of most mammalian miRNPs that mediates important functions in small RNA-directed regulatory pathways.
Argonaute (Ago) proteins are a family of proteins of ~95 kDa that bind directly to mature miRNAs. In mammals, there are four Ago proteins (Ago1-4). This family of proteins is a central component of most mammalian miRNPs that mediates important functions in small RNA-directed regulatory pathways.
Spécificité
This antibody recognizes Ago1, Ago2, Ago3, and Ago4. Note that this antibody has been reported to cross-react with radixin (~70 kDa). (Nelson, P., et al. (2007). RNA. 13:1787–1792.)
Immunogène
Histidine - tagged recombinant protein corresponding to human Ago.
Application
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
RNA Metabolism & Binding Proteins
Transcription Factors
RNA Metabolism & Binding Proteins
This RIPAb+ pan Ago -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Western Blot Analysis:
Representative lot data.
HeLa cell lysate was probed with Anti-pan Ago, clone 2A8 (0.5 μg/mL). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and chemiluminescence detection system.
Arrows indicates Ago (~ 95 kDa) and Radixin (~70 kDa). (Figure 2).
Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in IP. (Nelson, P., et al. (2007). RNA. 13:1787–1792.)
Immunocytochemistry Analysis: A representative lot was used by an independent laboratory in IC. (Nelson, P., et al. (2007). RNA. 13:1787–
1792.)
Immunohistochemistry Analysis: A representative lot was used by an independent laboratory in IH. (Nelson, P., et al. (2007). RNA. 13:1787–
1792.)
Representative lot data.
HeLa cell lysate was probed with Anti-pan Ago, clone 2A8 (0.5 μg/mL). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and chemiluminescence detection system.
Arrows indicates Ago (~ 95 kDa) and Radixin (~70 kDa). (Figure 2).
Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in IP. (Nelson, P., et al. (2007). RNA. 13:1787–1792.)
Immunocytochemistry Analysis: A representative lot was used by an independent laboratory in IC. (Nelson, P., et al. (2007). RNA. 13:1787–
1792.)
Immunohistochemistry Analysis: A representative lot was used by an independent laboratory in IH. (Nelson, P., et al. (2007). RNA. 13:1787–
1792.)
Conditionnement
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
Qualité
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 of Anti-pan Ago antibodyand the Magna RIP® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Argonaute proteins-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.
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 of Anti-pan Ago antibodyand the Magna RIP® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Argonaute proteins-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.
Description de la cible
~ 95 and 70 kDa observed. This antibody recognizes Ago1, Ago2, Ago3, and Ago4.
Note that this antibody has been reported to cross-react with radixin (~70 kDa). (Nelson, P., et al. (2007). RNA. 13:1787–1792.)
Note that this antibody has been reported to cross-react with radixin (~70 kDa). (Nelson, P., et al. (2007). RNA. 13:1787–1792.)
Forme physique
Anti-pan Ago (Mouse Monoclonal).One vial containing 50 μg of purified mouse IgG1κ in 0.1 M Tris-Glycine (pH 7.4) and 150 mM NaCl with 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C.
Normal Mouse IgG. One vial containing 125 µg of purified mouse IgG in 125 µL of 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
Normal Mouse IgG. One vial containing 125 µg of purified mouse IgG in 125 µL of 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
Format: Purified
Protein G
Stockage et stabilité
Stable for 1 year at -20°C 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: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
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: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Remarque sur l'analyse
Control
Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human IGF2 mRNA.
Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human IGF2 mRNA.
Autres remarques
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Informations légales
MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Clause de non-responsabilité
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.
Code de la classe de stockage
10-13 - German Storage Class 10 to 13
Certificats d'analyse (COA)
Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".
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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.
A high throughput experimental approach to identify miRNA targets in human cells.
Tan, Lu Ping, et al.
Nucleic Acids Research, 37, e137-e137 (2009)
Involvement of AGO1 and AGO2 in mammalian transcriptional silencing.
Janowski, BA; Huffman, KE; Schwartz, JC; Ram, R; Nordsell, R; Shames, DS; Minna, JD; Corey, DR
Nature Structural and Molecular Biology null
Mathieu Bertrand et al.
Developmental dynamics : an official publication of the American Association of Anatomists, 230(2), 325-334 (2004-05-27)
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