03-248
RIPAb+ pan Ago - RIP Validated Antibody and Primer Set
clone 2A8, from mouse
Synonym(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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All Photos(2)
About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32
Recommended Products
biological source
mouse
Quality Level
antibody form
purified immunoglobulin
clone
2A8, monoclonal
species reactivity
human
manufacturer/tradename
RIPAb+
Upstate®
technique(s)
RIP: suitable
immunocytochemistry: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG1κ
NCBI accession no.
UniProt accession no.
shipped in
dry ice
General description
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.
Specificity
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.)
Immunogen
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.)
Packaging
10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
Quality
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.
Target description
~ 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.)
Physical form
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
Storage and Stability
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.
Analysis Note
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.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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 Class Code
10-13 - German Storage Class 10 to 13
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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