RIPAb+ PUM2

Stock Code: 3586270
Manufacturer Part No: 03-241

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Descriptions

Analysis Note

ControlIncludes normal rabbit IgG and primers specific for human NFKBIA mRNA.

Application

Detect RIPAb+ PUM2 using this RIPAb+ PUM2 Antibody validated for use in ICC, IHC(P), Western Blotting, RNA Binding Protein Immunoprecipitation (RIP).

Western Blot Analysis:
Representative lot data.
Mouse embryonic stem cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-Pumilio 2 (PUM2) 1 µg/mL. Proteins were visualized using a Donkey Anti-Rabbit IgG conjugated to HRP and chemiluminescence detection system.
Arrow indicates PUM2 (~120 kDa).
Note: A non-specific band may be observed at ~48 kDa in some lysates. (Figure 2).
Immunohistochemistry (Parafin) Analysis:
Representative lot data.
Paraffin-embedded human testis (Fig. 3A) and human teratoma (Fig. 3B) tissues were prepared using heat-induced epitope retrieval in citrate buffer, pH 6.0. Immunostaining was performed using a 1:400 dilution of Anti-Pumilio 2 (PUM2). Reactivity was detected using the IHC-Select Detection Kit (Cat. No. DAB050). Staining pattern appears to be restricted to primary spermatocytes with no Sertoli cell involvement, as expected.
Immunocytochemistry Analysis:
Representative lot data.
Confocal fluorescent analysis of mouse and human embryonic stem cells using Anti-Pumilio 2 (PUM2) (Red). Actin filaments have been labeled with Alexa Fluor^® 488 dye - Phalloidin (Green). Nucleus is stained with DAPI (Blue). (Figure 4) This antibody positively stains the nucleus and cytosol.

Research CategoryStem Cell Research

Research Sub CategoryRNA Metabolism & Binding Proteins

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.
PUM2 is a sequence-specific RNA-binding protein that regulates translation and mRNA stability by binding to the 3′-UTR of mRNA targets. The PUM class of proteins plays an active role in the sustainability of mitotic stem cell proliferation and the regulation of cell development and differentiation.

Immunogen