RIPAb+ SMN - RIP Validated Antibody and Primer Set

The survival of motor neurons (SMN) protein is essential for the biogenesis of small nuclear RNA (snRNA)-ribonucleoproteins (snRNPs), the major components of the pre-mRNA splicing machinery. Though it is ubiquitously expressed, SMN deficiency causes the motor neuron degenerative disease spinal muscular atrophy (SMA). SMN deficiency has unexpected cell type-specific effects on the repertoire of snRNAs and mRNAs. It alters the stoichiometry of snRNAs and causes widespread pre-mRNA splicing defects in numerous transcripts of diverse genes, preferentially those containing a large number of introns, in SMN-deficient mouse tissues. The SMN complex plays a role in RNA metabolism and in splicing regulation.

Epitope: Unknown

Histidine-tagged recombinant protein corresponding to human SMN.

RNA Binding Protein Immunoprecipitation:
Representative lot data.
RIP Lysate prepared from HeLa cells (~2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using either 5 µg of a normal mouse IgG or 5 µg of Anti-SMN antibody and the Magna RIP^® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of SMN-associated RNA was verified by qPCR using primers specific for human collagen alpha type 1 mRNA (Please see figures).
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
HeLa cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with anti-SMN at a 0.5 µg/mL dilution.
Proteins were visualized using a goat anti-mouse IgG conjugated to HRP using a chemiluminescence detection system.
Arrow indicates SMN (~35 kDa) (Please see figures).

Immunocytochemistry Analysis:
Representative lot data.
A 1:500 dilution from a representative lot detected SMN in HeLa cells (Please see figures).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
RNA Metabolism & Binding Proteins

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

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

RNA Binding Protein Immunoprecipitation:
RIP Lysate prepared from HeLa cells (~2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using either 5 µg of a normal mouse IgG or 5 µg of Anti-SMN antibody and the Magna RIP^® RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
Successful immunoprecipitation of SMN-associated RNA was verified by qPCR using RIP Primers U1snRNA (Please see figures).
Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

~35 kDa was observed; however, the calculated molecular weight is 31.849 kDa

Anti-SMN (Mouse Monoclonal). One vial containing 50 µg of protein G purified monoclonal IgG1ĸ in buffer containing 0.1 M Tris-glycine, 150 mM NaCl, pH 7.4, 0.05% sodium azide before the addition of 30% glycerol. Store at -20°C.

Normal Mouse IgG. One vial containing 125 µg of purified mouse IgG in 125 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.

RIP Primers, U1snRNA. One vial containing 75 μL of 5 μM of each primer specific for the cDNA of U1 snRNP. Store at -20°C.
FOR: GGG AGA TAC CAT GAT CAC GAA GGT
REV: CCA CAA ATT ATG CAG TCG AGT TTC CC

Format: Purified

Protein G Purified

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. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Control
Includes negative control mouse IgG antibody and control primers specific for the cDNA of human U1snRNP.

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

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

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.