RIPAb+ Lin28 Kit

Stock Code: 3586251
Manufacturer Part No: 03-105
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Analysis Note


ControlIncluded negative control rabbit IgG antibody and control primers specific for CDK4.


Application


RNA Binding Protein Immunoprecipitation:
RIP Lysate prepared from NTera-2 embryonic carcinoma cells (2 X 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µl of either a normal rabbit IgG, or Anti-Lin28 antibody and the Magna RIP® Kit (Cat. # 17-700).
Successful immunoprecipitation of Lin28-associated RNA was verified by qPCR using primers for U1snRNA as a negative control and positive control RIP Primers CDK4 (Please see figures). Data is presented as percent input of each IP sample relative to input total RNA for each amplicon and RIP sample as indicated. Please refer to the Magna RIP™ (Cat. # 17-700) or EZ-Magna RIP™ (Cat. # 17-701) protocol for experimental details.

Immunoprecipitation from RIP lysate:
RIP lysate from NTera-2 embryonic carcinoma cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using 0.5 µL of either a normal rabbit IgG or Anti-Lin28 antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Lin28 (1.0 µg/mL). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00231) (Please see figures).


Research CategoryEpigenetics & Nuclear Function


Research Sub CategoryRNA Binding Protein (RBP)RNA Metabolism & Binding ProteinsSignaling Neuroscience


This RIPAb+ Lin28 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.


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 quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully coprecipitating a specific RNA target (where such a specific target is known). The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. Each set also includes a negative control antibody to ensure specificity of the RIP reaction.
The RIPAb+ Lin28 set includes the Lin28 (Zinc Finger CCHC domain-containing protein 1) antibody, a negative control antibody (purified rabbit IgG), and positive qPCR primers which amplify a 68 bp fragment of the human cDNA of CDK4. The Lin28 and negative control antibodies are supplied in a scalable "per RIP" reaction size and can be used to functionally validate the precipitation of Lin28-associated RNA.


Immunogen


The Lin28 antibody is made against full-length His-tagged human Lin28 recombinant protein.


Legal Information


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


Quality Level100, 500
ManufacturerSIGMA-ALDRICH

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