RIPAb+ CUGBP2 - RIP Validated Antibody and Primer Set

56 kDa

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.
CUGBP2 is a RNA-binding protein implicated in the regulation of several post-transcriptional events including pre-mRNA alternative splicing, mRNA translation and stability. CUGBP2 mediates exon inclusion and/or exclusion in pre-mRNA that are subject to tissue-specific and developmentally regulated alternative splicing.

A synthetic peptide corresponding to amino acids 209-415 of human CuGBP2.

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-CUGBP2 antibody (Cat. # CS204360).
Precipitated proteins (lane 1: mouse IgG, lane 2: anti-CUGBP2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-CUGBP2 antibody (Cat. # CS204360, 1:1000). Proteins were visualized using One-Step IP-Western kit (GenScript Cat. # L00231)
Arrow indicates CUGBP2 (Figure 2).
Western Blot Analysis:
Representative lot data.
Lysates from HL60 cells were probed with a previous lot of Anti-CUGBP2 (1:1000 dilution).
Arrow indicates CUGBP2 ~56 kDa (Figure 3).

Research Category
Protein Trafficking

Epigenetics & Nuclear Function

Research Sub Category
RNA Metabolism & Binding Proteins

RNA Binding Protein (RBP)

This RIPAb+ CUGBP2 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

CuGBP2

Other species have not yet been tested.

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

Anti-CUGBP2 (Mouse Monoclonal), Part # CS204360. One vial containing 50 µg of protein A purified mouse monoclonal IgG1 in 50 µL of 1X PBS, pH 7.0, and 0.1% 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

Format: Purified

Protein A purified

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.

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

RNA Binding Protein Immunoprecipitation:
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-CUGBP2 antibody (Cat. # CS204360) and the Magna RIP^® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of CUGBP2-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.

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

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

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

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.