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一般說明
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.
Ago3 (Eukaryotic translation initiation factor 2C 3) belongs to the argonaute family and contains a PAZ and a Piwi domain. Ago3 plays a role in the RNA interference (RNAi) pathway, which is important for heterochromatin formation and accurate chromosome segregation.
Ago3 (Eukaryotic translation initiation factor 2C 3) belongs to the argonaute family and contains a PAZ and a Piwi domain. Ago3 plays a role in the RNA interference (RNAi) pathway, which is important for heterochromatin formation and accurate chromosome segregation.
特異性
This antibody has been shown to not cross react with Ago1, Ago2, or Ago4 (Azuma-Mukai A et al. (2008) Proc. Natl. Acad. Sci. USA 105: 7964-7969).
免疫原
Recombinant protein corresponding to human Ago3
應用
Ago3 (Eukaryotic translation initiation factor 2C 3) belongs to the argonaute family & contains a PAZ & a Piwi domain. Ago3 plays a role in the RNA interference (RNAi) pathway.
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-Ago 3 antibody (Cat. # CS207366) and the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Ago 3-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.
Immunocytochemistry Analysis:
Representative lot data.
Confocal fluorescent analysis of HeLa cells using Anti-Ago3, clone 4B1-F6 (Red). Actin filaments have been labeled with Alexa Fluor 488 dye-Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
(Alexa Fluor is a registered trademark of Life Technologies.)
Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in HeLa cells. (Azuma-Mukai, A., et al. (2008). Proc Natl Acad Sci USA. 105(23):7964-7969.)
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-Ago 3 antibody (Cat. # CS207366) and the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Ago 3-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.
Immunocytochemistry Analysis:
Representative lot data.
Confocal fluorescent analysis of HeLa cells using Anti-Ago3, clone 4B1-F6 (Red). Actin filaments have been labeled with Alexa Fluor 488 dye-Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
(Alexa Fluor is a registered trademark of Life Technologies.)
Immunoprecipitation Analysis: A representative lot was used by an independent laboratory in HeLa cells. (Azuma-Mukai, A., et al. (2008). Proc Natl Acad Sci USA. 105(23):7964-7969.)
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
RNA Binding Protein (RBP)
RNA Binding Protein (RBP)
包裝
10 assays per set. Recommended use: ~5 µg of antibody per RIP (dependent upon biological context).
品質
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-Ago 3 antibody and the Magna RIP® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Argonaute 3-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 Anti-Ago 3 antibody and the Magna RIP® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of Argonaute 3-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.
標靶描述
~96 kDa
外觀
Protein G Purified
Anti-Ago3 (Mouse Monoclonal). One vial containing 50 µg of 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
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
Format: Purified
儲存和穩定性
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.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
分析報告
Control
Includes normal rabbit IgG and primers specific for human IGF2 mRNA.
Includes normal rabbit IgG and primers specific for human IGF2 mRNA.
其他說明
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
法律資訊
MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
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.
儲存類別代碼
10 - Combustible liquids
Dansen Wu et al.
Journal of cellular biochemistry, 116(12), 2744-2754 (2015-07-25)
Non-coding RNAs (ncRNAs) play key roles in epigenetic events. However, the exact mechanism of ncRNA guidance, particularly piwi-interacting RNAs (piRNAs), for the targeting of epigenetic regulatory factors to specific gene regions is unclear. Although piRNA function was first established in
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