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03-176

Sigma-Aldrich

RIPAb+ FXR1 - RIP Validated Antibody and Primer Set

from mouse

别名:

fragile X mental retardation, autosomal homolog 1, fragile X mental retardation-related protein 1

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About This Item

分類程式碼代碼:
12352203
eCl@ss:
32160702
NACRES:
NA.32

生物源

mouse

品質等級

抗體表格

purified immunoglobulin

無性繁殖

monoclonal

物種活性

mouse, human

製造商/商標名

RIPAb+
Upstate®

技術

RIP: suitable
immunocytochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

同型

IgG2aκ

NCBI登錄號

UniProt登錄號

運輸包裝

dry ice

基因資訊

human ... FXR1(8087)

一般說明

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.

特異性

This antibody recognizes FXR1.

免疫原

Recombinant protein corresponding to human FXR1.

應用

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).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
RNA Metabolism & Binding Proteins

Developmental Signaling
This RIPAb+ FXR1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

包裝

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

品質

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.

標靶描述

68 kDa

外觀

Protein G Purified
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
Format: Purified

儲存和穩定性

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.

分析報告

Control
Includes negative control mouse IgG antibody and control primers specific for the cDNA of human JUN.

其他說明

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

法律資訊

MAGNA RIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

免責聲明

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.

儲存類別代碼

10 - Combustible liquids


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Xianjian Wu et al.
Scientific reports, 14(1), 6155-6155 (2024-03-15)
As the most prevalent epitranscriptomic modification, N6-methyladenosine (m6A) shows important roles in a variety of diseases through regulating the processing, stability and translation of target RNAs. However, the potential contributions of m6A to RNA functions are unclear. Here, we identified
Shuo Qie et al.
Nature communications, 8(1), 1534-1534 (2017-11-17)
The Fbxo4 tumour suppressor is a component of an Skp1-Cul1-F-box E3 ligase for which two substrates are known. Here we show purification of SCFFbxo4 complexes results in the identification of fragile X protein family (FMRP, Fxr1 and Fxr2) as binding
Arantxa Agote-Aran et al.
The EMBO journal, 39(20), e104467-e104467 (2020-07-25)
Nucleoporins (Nups) build highly organized nuclear pore complexes (NPCs) at the nuclear envelope (NE). Several Nups assemble into a sieve-like hydrogel within the central channel of the NPCs. In the cytoplasm, the soluble Nups exist, but how their assembly is
Arantxa Agote-Arán et al.
Frontiers in cell and developmental biology, 9, 755847-755847 (2022-01-04)
Nuclear pore complexes (NPCs) are embedded in the nuclear envelope (NE) where they ensure the transport of macromolecules between the nucleus and the cytoplasm. NPCs are built from nucleoporins (Nups) through a sequential assembly order taking place at two different
Yongrong Liao et al.
The Journal of cell biology, 223(7) (2024-04-23)
Assembly of macromolecular complexes at correct cellular sites is crucial for cell function. Nuclear pore complexes (NPCs) are large cylindrical assemblies with eightfold rotational symmetry, built through hierarchical binding of nucleoporins (Nups) forming distinct subcomplexes. Here, we uncover a role

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