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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M | RIP, WB, ICC, IP |
| Description | |
|---|---|
| Catalogue Number | 03-176 |
| Trade Name |
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| Description | RIPAb+™ FXR1 - 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. Fragile X mental retardation-related protein 1 (FXR1) and FXR2 are members of the FMR1 family sharing a very high degree of protein similarity among them. FXR1 is an RNA binding protein and is expressed in the heart, kidney, brain, and testis. Mutations in FMR1 result in the most common form of mental retardation. |
| Alternate Names |
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| Background Information | Fragile X mental retardation-related protein 1 (FXR1) and FXR2 are members of the FMR1 family sharing a very high degree of protein similarity among them. They interact with each other and also with FMR1, moving between the cytoplasm and the nucleus. FXR1 is an RNA binding protein and is expressed in the heart, kidney, brain, and testis. Mutations in FMR1 result in the most common form of mental retardation. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Anti-FXR1 (Mouse Monoclonal), Part # CS204403. One vial containing 50 µg of protein G purified monoclonal antibody in buffer containing 0.1 M Tris-glycine, 0.105 M NaCl, pH 7.4, 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C. Normal Mouse IgG, Part # CS200621. One vial containing 125 µg purified mouse IgG in 125 µL storage buffer containing 0.1% sodium azide. Store at -20°C. RIP Primers, JUN, Part # CS203198. One vial containing 75 μL of 5 μM of each primer specific for the cDNA of human JUN. Store at -20°C. FOR: TCG ACA TGG AGT CCC AGG A REV: GGC GAT TCT CTC CAG CTT CC |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This RIPAb+ FXR1 -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 mouse IgG, (Cat. #CS200621), or 5 µg of Anti-FXR1 antibody (Cat. # CS204403). Ten percent of the precipitated proteins (lane 1: T7, lane 2: mouse IgG, lane 4: FXR1) and HeLa whole cell lysate (lane 3) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-FXR1 antibody (Cat. # CS204403, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232). Arrow indicates Aly/REF (~68 kDa) (Figure 2). Western Blot Analysis: Representative lot data. HeLa cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with anti-FXR1 (0.5 μg/mL) dilution. Proteins were visualized using a Goat anti-mouse IgG conjugated to HRP using a chemiluminescence detection system. Arrow indicates FXR1 (~68 kDa) (Figure 3). Immunocytochemistry Analysis: Representative lot data. A 1:500 dilution from a representative lot detected FXR1 in HeLa and NIH/3T3 cell lysates. Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytoplasm (Figure 4). Immunoprecipitation Analysis: A representative lot was used in an independent laboratory in immunoprecipitation. (Siomi, M., et al. (1996). Molecular and Cellular Biology. 3825-3832). |
| Biological Information | |
|---|---|
| Immunogen | Recombinant protein corresponding to human FXR1. |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | This antibody recognizes FXR1. |
| Isotype | IgG2aκ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The protein encoded by this gene is an RNA binding protein that interacts with the functionally-similar proteins FMR1 and FXR2. These proteins shuttle between the nucleus and cytoplasm and associate with polyribosomes, predominantly with the 60S ribosomal subunit. Three transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq] |
| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: RNA-binding protein required for embryonic and postnatal development of muscle tissue. May regulate intracellular transport and local translation of certain mRNAs By similarity. SUBUNIT STRUCTURE: Interacts with FMR1 and FXR2. Interacts with TDRD3, and with CYFIP2 but not with CYFIP1. SUBCELLULAR LOCATION: Cytoplasm. TISSUE SPECIFICTY: Expressed in all tissues examined including heart, brain, kidney and testis. PTM: Arg-445 is dimethylated, probably to asymmetric dimethylarginine. SEQUENCE SIMILARITIES: Belongs to the FMR1 family. Contains 2 KH domains. |
| Molecular Weight | 68 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | RNA Binding Protein Immunoprecipitation: RIP Lysate prepared from Jurkat cells (~5 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using either 5 µg of a normal mouse IgG (Cat. # CS200621), or 5 µg of Anti-FXR1 antibody (Cat. # CS204403) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of Aly/REF-associated RNA was verified by qPCR using RIP Primers JUN, (Cat. # CS203198) (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: Variability 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). |