RIPAb+ IGF2 mRNA-binding protein 3 - RIP Validated Antibody and Primer Set

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.
IGF2 mRNA-binding protein 3 contains several KH domains, which are important in RNA binding and are known to be involved in RNA synthesis and metabolism.

This antibody recognizes IGF2 mRNA-binding protein 3 at the C-terminus.

Epitope: C-terminus

KLH-conjugated linear peptide corresponding to human IGF2 mRNA-binding protein 3 at and around the C-terminus.

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-IGF2 mRNA-binding protein 3 antibody (Cat. # CS207310). Ten percent of the precipitated proteins (lane 1: Rabbit IgG, lane 2: IGF2 mRNA-binding protein 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-IGF2 mRNA-binding protein 3 antibody (Cat. # CS207310, 1:1000). Proteins were visualized using One-Step IP-Western kit (GenScript Cat. # L00231) Arrow indicates IGF2 mRNA-binding protein 3. The lower band in lane 2 correspond to the isoform 2.
Western Blot Analysis:
Representative lot data.
HEK293 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with IGF2BP3, (1:2,000) dilution. Proteins were visualized using a Donkey anti-Rabbit conjugated to HRP using a chemiluminescence detection system.
Arrow indicates IGF2BP3 (~64 kDa).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Growth Factors & Receptors

RNA Metabolism & Binding Proteins

This RIPAb+ IGF2 mRNA-binding protein 3 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).

RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from HepG2 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-IGF2 mRNA-binding protein antibody and the Magna RIP^® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of IGF2 mRNA-binding protein 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.

64 kDa was observed. Predicted to cross-react with all the Isoforms of IGF2BP3.

Affinity purified

Anti-IGF2 mRNA-binding protein 3 (Rabbit Polyclonal). One vial containing 75 μL of affinity purified rabbit polyclonal antibody in PBS with 0.09% sodium azide and 30% glycerol. Store at -20°C. Concentration: 0.7 mg/mL
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 Primer IGF2, Part # CS207311.
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

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.

Control
Includes negative control normal rabbit IgG antibody and control primers specific for the cDNA of human IGF2.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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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.