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  • 03-178 - RIPAb+ SMN - RIP Validated Antibody and Primer Set

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03-178 Sigma-Aldrich

RIPAb+ SMN - RIP Validated Antibody and Primer Set

from mouse

Synonym: Component of gems 1, gemin 1, spinal muscular atrophy (Werdnig-Hoffmann disease, Kugelberg-Welander disease), survival of motor neuron 1, telomeric

  •  eCl@ss 32160702

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Properties

Related Categories Antibodies, Antibody Kits, Immunoprecipitation Kits More...
clone   monoclonal
biological source   mouse
application(s)   RIP: suitable
  immunocytochemistry: suitable
  immunoprecipitation (IP): suitable
  western blot: suitable
species reactivity   Xenopus, mouse, human, human
species reactivity (predicted by homology)   Xenopus, mouse
shipped in   dry ice
isotype   IgG1κ
Quality Level   100
mfr. no.   RIPAb+
  Upstate®
NCBI accession no.   NP_000335.1
UniProt accession no.   Q16637
Gene Information   human ... SMA4(11039)

Description

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.

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.

Immunogen

Epitope: Unknown

Histidine-tagged recombinant protein corresponding to human SMN.

Application

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.

Target description

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

Physical form

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

Storage and Stability

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.

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.

Legal Information

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

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

Packaging

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

Quality

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.

Analysis Note

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

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

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Peer-Reviewed Papers
15

References

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