RIPAb+ GFP - RIP Validated Antibody

RNA Binding Protein Immunoprecipitation:
RIP Lysate prepared from HeLa cells (1 X 10E7 cell equivalents per IP) expressing SNRNP70-GFP-FLAG (Cat. No. 03-903) by RNA transfection was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or Anti-Green Fluorescent Protein Antibody and the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of SNRNP70-GFP-FLAG associated RNA was verified by qPCR using RIP Primers, U1 snRNA.

Immunoprecipitation:
RIP Lysate prepared from HeLa cells (1 X 10E6 cell equivalents per IP) expressing SNRNP70-GFP-FLAG (Cat. No. 03-903) by RNA transfection was subjected to immunoprecipitation using 0.5 µg of either a normal rabbit IgG, or anti-Green Fluorescent Protein Antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Green Fluorescent Protein Antibody (1.0 µg/mL). Proteins were visualized using Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP (Cat. # AP187P) and a chemiluminescence detection system.

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

The anti-Green Fluorescent Protein Antibody is made against highly-purified native GFP from Aequorea victoria. It is reactive with GFP from both native and recombinant sources. While other commercial antibodies to GFP typically demonstrate strong reactivity with E. coli, the antibody has been purified by Protein A chromatography, followed by extensive adsorption against whole E. coli BL21 strain cells, giving them minimal cross-reactivity with E. coli.

Highly purified native GFP from Aequorea victoria

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, 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

Anti-Green Fluorescent Protein Antibody: One vial containing 50 μg of protein G purified antibody in 50 μL buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4). Store at -20°C. Normal Mouse IgG: One vial containing 125 μg purified mouse IgG in 125 μL storage buffer containing 0.1% sodium azide. Store at -20°C. RIP Primers, U1 snRNA : One vial containing 75 μL of 5 μM of each primer specific for the cDNA of human U1 snRNA. Store at -20°C.FOR: GGG AGA TAC CAT GAT CAC GAA GGTREV: CCA CAA ATT ATG CAG TCG AGT TTC CC

RNA Binding Protein Immunoprecipitation: RIP Lysate prepared from HeLa cells (1 X 10E7 cell equivalents per IP) expressing SNRNP70-GFP-FLAG (Cat. No. 03-903) by RNA transfection was subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or Anti-Green Fluorescent Protein Antibody and the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of SNRNP70-GFP-FLAG associated RNA was verified by qPCR using RIP Primers, U1 snRNA.
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

The Green Fluorescent Protein (GFP) from the jellyfish Aequorea victoria is used as a fluorescent indicator for monitoring gene expression in a variety of cellular systems, including living organisms and fixed tissues. Unlike other bioluminescent reporters, GFP fluoresces in the absence of substrates, cofactors, or other intrinsic or extrinsic proteins. Purified GFP is a 27 kDa monomer consisting of 238 amino acids and emits green light (emission maximum at 509 nm) when excited with blue or UV light.

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.