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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M, Chp, Ca, Eq | RIP, WB, IP |
| Description | |
|---|---|
| Catalogue Number | 03-198 |
| Trade Name |
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| Description | RIPAb+™ IGF2 mRNA-binding protein 3 - RIP Validated Antibody and Primer Set |
| 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. 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. |
| Alternate Names |
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| Background Information | The protein encoded by this gene is primarily found in the nucleolus, where it can bind to the 5 UTR of the insulin-like growth factor 2 leader 3 mRNA and may repress translation of insulin-like growth factor 2 during late development. The encoded protein contains several KH domains, which are important in RNA binding and are known to be involved in RNA synthesis and metabolism. A pseudogene exists on chromosome 7, and there are putative pseudogenes on other chromosomes. [provided by RefSeq]. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
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| Presentation | 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 |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This RIPAb+ IGF2 mRNA-binding protein 3 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
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| Application Notes | 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). |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to human IGF2 mRNA-binding protein 3 at and around the C-terminus. |
| Epitope | C-terminus |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This antibody recognizes IGF2 mRNA-binding protein 3 at the C-terminus. |
| Species Reactivity |
|
| Species Reactivity Note | Reacts with human and mouse. Predicted to react with chimpanzee, Rhesus monkey, dog, and horse based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The protein encoded by this gene is primarily found in the nucleolus, where it can bind to the 5 UTR of the insulin-like growth factor 2 leader 3 mRNA and may repress translation of insulin-like growth factor 2 during late development. The encoded protein contains several KH domains, which are important in RNA binding and are known to be involved in RNA synthesis and metabolism. A pseudogene exists on chromosome 7, and there are putative pseudogenes on other chromosomes. [provided by RefSeq]. |
| Gene Symbol |
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| Purification Method | Affinity Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: RNA-binding protein that act as a regulator of mRNA translation and stability. Binds to the 5-UTR of the insulin-like growth factor 2 (IGF2) mRNAs. Binds to sequences in the 3-UTR of CD44 mRNA. SUBUNIT STRUCTURE: Homodimer and multimer. SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note: Found in lamellipodia of the leading edge, in the perinuclear region, and beneath the plasma membrane. The subcytoplasmic localization is cell specific and regulated by cell contact and growth. Localized at the connecting piece and the tail of the spermatozoa. Colocalized with CD44 mRNA in RNP granules. TISSUE SPECIFICITY: Expressed in fetal liver, fetal lung, fetal kidney, fetal thymus, fetal placenta, fetal follicles of ovary and gonocytes of testis, growing oocytes, spermatogonia and semen (at protein level). Expressed in cervix adenocarcinoma, in testicular, pancreatic and renal-cell carcinomas (at protein level). Expressed ubiquitously during fetal development at 8 and 14 weeks of gestation. Expressed in ovary, testis, brain, placenta, pancreatic cancer tissues and pancreatic cancer cell lines. INVOLVEMENT IN DISEASE: Autoantibodies against IGF2BP3 are detected in sera from some patients with a variety of carcinomas. SEQUENCE SIMILARITIES: Belongs to the RRM IMP/VICKZ family. Contains 4 KH domains. Contains 2 RRM (RNA recognition motif) domains. |
| Molecular Weight | 64 kDa was observed. Predicted to cross-react with all the Isoforms of IGF2BP3. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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. |
| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | 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. |
| Packaging Information | |
|---|---|
| Material Size | 10 assays |
| Material Package | 10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context). |