Analysis Note

ControlIncludes negative control normal rabbit IgG antibody and control primers specific for the human ARF1 mRNA.

Application

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryTranscription FactorsRNA Metabolism & Binding Proteins

Immunoprecipitation from RIP lysate:
Representative lot data.
RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal rabbit IgG, (Cat. #PP64B), or 5 µg of Anti-STAU1 (Staufen 1) antibody (Cat. # CS204405). Ten percent of the precipitated proteins (lane 1: rabbit IgG, lane 2: STAU1) were resolved by electrophoresis, transferred to nitrocellulose and probed with Anti-STAU1 (Staufen 1) antibody (Cat. # CS204405, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) .
Arrow indicates STAU1. (Figure 2).
Western Blot Analysis:
Representative lot data.
HepG2 cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-STAU1 antibody (Cat. # CS204405, 1:1000 dilution). Proteins were visualized using a Goat Anti-Rabbit IgG conjugated to HRP and a chemiluminescence detection system.
Arrow indicates STAU1 (~63 kDa). (Figure 3).
Note: Uncharacterized bands appear at ~22 kDa and ~53 kDa in

Disclaimer

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.

General description

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.
STAU1 (Staufen 1) is a RNA-binding protein implicated in the transport and/or localization of mRNA via the microtubule network to the rough endoplasmic reticulum.

Immunogen

Epitope: a.a. 2-51

Synthetic peptide from amino acids 2-51 of human STAU 1.

Legal Information

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

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

Packaging

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

Physical form

Anti-STAU1 (Staufen 1) (Rabbit Polyclonal),. Lyophilized powder. Add 50 µL of distilled water. Final antibody concentration is 1 mg/ml in PBS buffer. For longer periods of storage, store at -20°C. Avoid repeat freeze-thaw cycles.
Normal Rabbit IgG. One vial containing 125 µg of purified rabbit IgG in 125 µL of storage buffer containing 0.05% sodium azide. Store at -20°C.
RIP Primers, ARF1.
One vial containing 75 µL of 5 µM of each primer specific for human ARF1 mRNA.
Store at -20°C.
FOR: GAC CAC GAT CCT CTA CAA GC
REV: TCC CAC ACA GTG AAG CTG ATG

Format: Purified

Quality

RNA Binding Protein Immunoprecipitation:
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal Rabbit IgGor 5 µg of Anti-STAU1 antibody and the Magna RIP^® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of STAU1-associated RNA was verified by qPCR using RIP Primers ARF1, (Figure 1).
Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.

Specificity

Other species have not yet been tested.

Storage and Stability

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.

Target description

~63 kDa observed. Uncharacterized bands appear at ~22 kDa and ~53 kDa in some lysates.