In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures.
The principle of RNA EMSA as well as DNA EMSA is simple, considerations for performing a successful RNA EMA including the follows:
1）Needs instruments and reagents for RNA EMSA.
2) The knowledge for handling RNAs and proteins.
3) The skills for running a vertical gel electrophoresis.
4) The facility for imaging ECL signals via x-ray films or imagers for imaging ECL or infrared fluorescence.
5) The RNA probe is expensive, 20 fold of what for making a DNA probes.
6) Most probes for RNA EMSA is a single-strand RNA, which is very fragile for degradation.
7) Working environment should keep as RNase-free.
8) Based on the considerations above, we recommend our IRFluo rEMSA kits for RNA EMSA for its fast results (~2 hours) and simple operation (imaging gels immediately after gel electrophoresis).
9) If using Viagene's RNA EMSA service, the results would be secured as the scientists in Viagene Biotech have extensive experience for RNA EMSA (Linking to technique services).
|SIDET101||Coolshift-BId||General ECL EMSA kit, DNA||100assays||277|
|SIDET102||Coolshift-BIr||General ECL EMSA kit, RNA||100assays||399|
|SIDET201||Coolshift-IRd||General IR EMSA kit, DNA||100assays||199|
|SIDET202||Coolshift-IRr||General IR EMSA kit, RNA||100assays||298|
|BITFxxx||detail||Complete ECL EMSA kits||30/60assays||287/492|
|IRTFxxx||detail||Complete IR EMSA kits||30/60assays||287/492|
|IRTFxxx||detail||Gel-staining EMSA kits||50/100assays||129/198|
|SERV005a1||EMSA-PN Ctrl||EMSA/positive+negatives||5 loads/ea.||95|
|SERV005a2||EMSA-PN Ctrl||EMSA/positive+negatives||10 loads/ea.||155|
|SERV005b1||EMSA-P Ctrl||EMSA/positive controls||5 loads/ea.||75|
|SERV005b2||EMSA-P Ctrl||EMSA/positive controls||10 loads/ea.||119|
|SERV005c1||EMSA-N Ctrl||EMSA/negative controls||5 loads/ea.||45|
|SERV005c2||EMSA-N Ctrl||EMSA/negative controls||10 loads/ea.||59|
|TFRGT103a||EMSA-BBuf||ECL EMSA/blocking buf.||60ml/btl||45|
|TFRGT103b||EMSA-WBuf||ECL EMSA/washing buf.||60ml/btl||59|
|TFRGT103c||EMSA-EBuf||ECL EMSA/equilibration buf.||60ml/btl||21|
|TFxxxBP||details||Biotin-probes for EMSA||50assays||99|
|TFxxxBP||details||Biotin-probes for EMSA||100assys||159|
|TFxxxIR||details||IR Fluo-probes for EMSA||50assys||135|
|TFxxxIR||details||IR Fluo-probes for EMSA||100assys||199|
|TFxxxCP||details||Unlabeled competitive probes||6assays||45|
|TFxxxCP||details||Unlabeled competitive probes||12assays||65|
|TFxxxMP||details||Unlabeled mutant probes||6assays||59|
|TFxxxMP||details||Unlabeled mutant probes||12assays||89|
|SINP001a||C-N extractn||Cell Nuclear Extraction||50samples||99|
|SINP001b||T-N extractn||Tissue Nuclear Extraction||50samples||125|
|SINP013||EMSA-ProC||Nucl. protein concentration||100mL/botl||85|