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03-108
Sigma-AldrichRIPAb+ Fragile X Mental Retardation Protein - RIP Validated Antibody and Primer Set
This RIPAb+ Fragile X Mental Retardation Protein -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This RIPAb+ Fragile X Mental Retardation Protein -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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RIPAb+ Fragile X Mental Retardation Protein - 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.
Alternate Names
Fragile X mental retardation 1 protein
Background Information
Fragile X Mental Retardation Protein (FMRP) is a RNA-binding protein that is associated to polysomes and may be involved in the transport of mRNA from the nucleus to the cytoplasm. Defects in FMR1 are the cause of Fragile X syndrome, which is a common genetic disease characterized by moderate to severe mental retardation, macroorchidism, large ears, prominent jaw, and high-pitched, jocular speech. The defect in most fragile X syndrome patients results from an amplification of a CGG repeat region which is directly in front of the coding region.
References
Product Information
Format
Purified
Control
Includes negative control mouse IgG antibody and control primers specific for mouse Iqsec1.
Presentation
Anti-Fragile X Mental Retardation Protein (Mouse Monoclonal IgG1κ). One vial containing 50 μL of liquid, does not contain any preservative. 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 Primers, Iqsec1.
One vial containing 75 μL of 5 μM of each primer specific for mouse Iqsec1. Store at -20°C. FOR: GCG TCT AGG ACC TTG TGA GC REV: GAA GGC TGA CCT CCA TGT TG
This RIPAb+ Fragile X Mental Retardation Protein -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
RNA Binding Protein Immunoprecipitation (RIP)
Western Blotting
ELISA
Immunocytochemistry
Immunohistochemistry
Immunohistochemistry (Paraffin)
Immunoprecipitation
Application Notes
Western Blot Analysis: Representative lot data. Mouse E17 spinal cord lysate (Lane 1), Mouse E16 brain (Lane 2) and NIH/3T3 lysate (Lane 3) were resolved by electrophoresis, transferred to PVDF membrane and probed with anti-Fragile X Mental Retardation Protein (1:500 dilution of a previous lot). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates Fragile X Mental Retardation Protein (~71 kDa) (Please see figures).
Immunohistochemistry: Frozen and paraffin sections. 1:500-1:5,000 dilution of a previous lot was used.
ELISA: A 1:500-1:5,000 dilution of a previous lot was used in ELISA.
Detection of FMRP on Blood Smears: A 1:500-1:5,000 dilution of a previous lot was used.
Immunocytochemistry: A 1:500-1:5,000 dilution of a previous lot was used on transfected cells. Light fixation (2% PFA, permeabilize with 0.1% triton in block only) (Please see figures).
Optimal working dilutions must be determined by the end user.
Biological Information
Immunogen
Fusion protein with a full length FMRP (human).
Host
Mouse
Specificity
FMRP. Cross reaction with FXR protein may be detected in cases of high expression in mice.
The protein encoded by this gene binds RNA and is associated with polysomes. The encoded protein may be involved in mRNA trafficking from the nucleus to the cytoplasm. A trinucleotide repeat (CGG) in the 5' UTR is normally found at 6-53 copies, but an expansion to 55-230 repeats is the cause of fragile X syndrome.
FUNCTION: SwissProt: Q06787 # RNA-binding protein. Associated with polysomes and might be involved in the transport of mRNA from the nucleus to the cytoplasm. Binds strongly to poly(G), binds moderately to poly(U) but shows very little binding to poly(A) or poly(C). SIZE: 632 amino acids; 71174 Da SUBUNIT: Found in a RNP granule complex with IGF2BP1. Interacts with FXR1, FXR2, IGF2BP1, NUFIP1 and RANBP9. Interacts with CYFIP1 and CYFIP2. SUBCELLULAR LOCATION: Cytoplasm. Nucleus. TISSUE SPECIFICITY: Highest levels found in neurons, brain, testis, placenta and lymphocytes. Also expressed in epithelial tissues and at very low levels in glial cells. DISEASE: SwissProt: Q06787 # Defects in FMR1 are the cause of fragile X syndrome. [MIM:300624]. It is a common genetic disease (has a prevalence of one in every 2000 children) which is characterized by moderate to severe mental retardation, macroorchidism (enlargement of the testicles), large ears, prominent jaw, and high-pitched, jocular speech. The defect in most fragile X syndrome patients results from an amplification of a CGG repeat region which is directly in front of the coding region. & Defects in FMR1 are the cause of fragile X tremor/ataxia syndrome (FXTAS) [MIM:300623]. In FXTAS, the expanded repeats range in size from 55 to 200 repeats and are referred to as 'premutations'. Full repeat expansions with greater than 200 repeats results in fragile X mental retardation syndrome [MIM:300624]. Carriers of the premutation typically do not show the full fragile X syndrome phenotype, but comprise a subgroup that may have some physical features of fragile X syndrome or mild cognitive and emotional problems. SIMILARITY: SwissProt: Q06787 ## Belongs to the FMR1 family. & Contains 2 KH domains. MISCELLANEOUS: RNA-binding activity is inhibited by RANBP9. & The mechanism of the severe phenotype in the Asn- 304 patient lies in the sequestration of bound mRNAs in nontranslatable mRNP particles. In the absence of FMRP, these same mRNAs may be partially translated via alternate mRNPs, although perhaps abnormally localized or regulated, resulting in typical fragile X syndrome. Asn-304 mutation maps to a position within the second KH domain of FMRP that is critical for stabilizing sequence-specific RNA-protein interactions. Asn-304 mutation abrogates the association of the FMRP KH 2 domain with its target, kissing complex RNA.
Molecular Weight
71 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
RNA Binding Protein Immunoprecipitation: RIP Lysate prepared from NIH/3T3 cells (2 x 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µL of either a normal mouse IgG or 5 µL of Anti-Fragile X Mental Retardation Protein antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of Fragile X Mental Retardation Protein-associated RNA was verified by qPCR using RIP Primers Iqsec1 (Please see figures). Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.
Usage Statement
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 and Shipping Information
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.
Packaging Information
Material Size
10 assays
Material Package
10 assays per set. Recommended use: ~5 μL of antibody per RIP (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
03-108
04053252679193
Documentation
RIPAb+ Fragile X Mental Retardation Protein - RIP Validated Antibody and Primer Set SDB
