RIPAb+™ FXR2 - 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.
Regulates transcription by acting as a general inhibitor of RNA polymerase II. When HEXIM1 is knocked down, HEXIM2 functionally compensates for its association with P-TEFb. HEXIM2 is up-regulated by HMBA (hexamethylene bisacetamide) at protein level.
Background Information
Fragile X Mental Retardation Syndrome is the most common form of hereditary mental retardation, and is caused by defects in the FMR1 gene. FMR1 is an RNA-binding protein and the syndrome results from lack of expression of FMR1 or expression of a mutant protein that is impaired in RNA binding. FXR2 is very similar to FMR1 (60% identity). FXR2 encodes a 74 kDa protein which, like FMR1, contains two KH domains, has the capacity to bind RNA and is localized to the cytoplasm. The FXR2 gene is located on human chromosome 17 at 17p13.1. In addition, FMR1 and FXR2 interact tightly with the recently described autosomal homolog FXR1. Each of these three proteins is capable of forming heterodimers with the others, and each can also form homodimers. FXR1 and FXR2 are thus likely to play important roles in the function of FMR1 and in the pathogenesis of the Fragile X Mental Retardation Syndrome.
Product Information
Format
Purified
Control
Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human JUN.
Presentation
Anti-FXR2 (Mouse Monoclonal), Part # CS207315. 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 of purified mouse IgG in 125 µL of 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
Applications
Application
This RIPAb+ FXR2 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
RNA Binding Protein Immunoprecipitation (RIP)
Application Notes
Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from Jurkat cells (~5 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-FXR2 antibody (Cat. # CS207315). Twenty percent of the precipitated proteins (lane 1: mouse IgG, lane 2: FXR2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-FXR2 antibody (Cat. # CS207315, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) Arrow indicates FXR2 (~74 kDa). (Figure 2).
Western Blot Analysis:
Representative lot data.
Hek293 cell lysate was probed with Anti-FXR2 (0.5 µg/mL). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates FXR2 (~74kDa). (Figure 3).
Biological Information
Immunogen
Recombinant protein corresponding to human FXR2.
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
The protein encoded by this gene is a RNA binding protein containing two KH domains and one RCG box, which is similar to FMRP and FXR1. It associates with polyribosomes, predominantly with 60S large ribosomal subunits. This encoded protein may self-associate or interact with FMRP and FXR1. It may have a role in the development of fragile X mental retardation syndrome. [provided by RefSeq].
SUBUNIT STRUCTURE: Interacts with FMR1 and FXR1. Interacts with CYFIP2 but not with CYFIP1. Interacts with TDRD3. Ref.9
SUBCELLULAR LOCATION: Cytoplasm.
SEQUENCE SIMILARITIES: Belongs to the FMR1 family.
Contains 2 KH domains.
Product Usage Statements
Quality Assurance
RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from Jurkat cells (~5 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG (Cat. # CS200621), or 5 µg of Anti-FXR2 antibody (Cat. # CS207315) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of FXR2-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
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. 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).
原厂资料：
Key Spec Table
Species Reactivity
Key Applications
M, H, R
WB, RIP
Description
Catalogue Number
03-246
Trade Name
Upstate
RIPAb+
Description
RIPAb+™ FXR2 - 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.
Regulates transcription by acting as a general inhibitor of RNA polymerase II. When HEXIM1 is knocked down, HEXIM2 functionally compensates for its association with P-TEFb. HEXIM2 is up-regulated by HMBA (hexamethylene bisacetamide) at protein level.
Background Information
Fragile X Mental Retardation Syndrome is the most common form of hereditary mental retardation, and is caused by defects in the FMR1 gene. FMR1 is an RNA-binding protein and the syndrome results from lack of expression of FMR1 or expression of a mutant protein that is impaired in RNA binding. FXR2 is very similar to FMR1 (60% identity). FXR2 encodes a 74 kDa protein which, like FMR1, contains two KH domains, has the capacity to bind RNA and is localized to the cytoplasm. The FXR2 gene is located on human chromosome 17 at 17p13.1. In addition, FMR1 and FXR2 interact tightly with the recently described autosomal homolog FXR1. Each of these three proteins is capable of forming heterodimers with the others, and each can also form homodimers. FXR1 and FXR2 are thus likely to play important roles in the function of FMR1 and in the pathogenesis of the Fragile X Mental Retardation Syndrome.
Product Information
Format
Purified
Control
Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human JUN.
Presentation
Anti-FXR2 (Mouse Monoclonal), Part # CS207315. 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 of purified mouse IgG in 125 µL of 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
Applications
Application
This RIPAb+ FXR2 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
RNA Binding Protein Immunoprecipitation (RIP)
Application Notes
Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from Jurkat cells (~5 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-FXR2 antibody (Cat. # CS207315). Twenty percent of the precipitated proteins (lane 1: mouse IgG, lane 2: FXR2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-FXR2 antibody (Cat. # CS207315, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) Arrow indicates FXR2 (~74 kDa). (Figure 2).
Western Blot Analysis:
Representative lot data.
Hek293 cell lysate was probed with Anti-FXR2 (0.5 µg/mL). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates FXR2 (~74kDa). (Figure 3).
Biological Information
Immunogen
Recombinant protein corresponding to human FXR2.
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
The protein encoded by this gene is a RNA binding protein containing two KH domains and one RCG box, which is similar to FMRP and FXR1. It associates with polyribosomes, predominantly with 60S large ribosomal subunits. This encoded protein may self-associate or interact with FMRP and FXR1. It may have a role in the development of fragile X mental retardation syndrome. [provided by RefSeq].
SUBUNIT STRUCTURE: Interacts with FMR1 and FXR1. Interacts with CYFIP2 but not with CYFIP1. Interacts with TDRD3. Ref.9
SUBCELLULAR LOCATION: Cytoplasm.
SEQUENCE SIMILARITIES: Belongs to the FMR1 family.
Contains 2 KH domains.
Product Usage Statements
Quality Assurance
RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from Jurkat cells (~5 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG (Cat. # CS200621), or 5 µg of Anti-FXR2 antibody (Cat. # CS207315) and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of FXR2-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
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. 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).
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