03-203 | RIPAb+ Gemin6 - RIP Validated Antibody and Primer Set

03-203
10 assays  10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey Applications
      HRIP, WB, ICC, IP
      Description
      Catalogue Number03-203
      Brand Family Upstate
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+ Gemin6 - RIP Validated Antibody and Primer Set
      OverviewRIPAb+ 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.
      Gemin6 is part of a large macromolecular complex that plays a role in the cytoplasmic assembly of small nuclear ribonucleoproteins (snRNPs). Other members of this complex include SMN, Gemin 2, Gemin 3, Gemin 4, and Gemin 5.
      Alternate Names
      • gem (nuclear organelle) associated protein 6
      • gemin 6
      Background InformationThe survival of motor neurons (SMN) complex is essential for the biogenesis of small nuclear ribonucleoprotein (snRNP) complexes in eukaryotic cells. Reduced levels of SMN cause the motor neuron degenerative disease, spinal muscular atrophy. Complexes containing Gemin3-Gemin4-Gemin5 and Gemin6-Gemin7-unrip persist at similar levels when SMN is reduced. Gemin5-containing subunits bind small nuclear RNA independently of the SMN complex and without a requirement for exogenous ATP. ATP hydrolysis is, however, required for displacement of small nuclear RNAs from the Gemin5-containing subunits and their assembly into snRNPs.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control normal mouse IgG antibody and control primers specific for the cDNA of human U1snRNA.
      PresentationAnti-Gemin6 (Mouse Monoclonal). One vial containing 50 μg of purified mouse IgG1k in buffer containing 0.1M Tris-Glycine (pH 7.4, 150 mM NaCl) with 0.05% sodium azide and 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
      Applications
      ApplicationThis RIPAb+ Gemin6 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • RNA Binding Protein Immunoprecipitation (RIP)
      • Western Blotting
      • Immunocytochemistry
      • Immunoprecipitation
      Application NotesImmunoprecipitation from RIP lysate:
      Representative lot data.
      RIP lysate from HeLa cells (~2 X 10E7 cell equivalents per IP) was subjected to immunoprecipitation using either 5 µg of a normal mouse IgG or 5 µg of Anti-Gemin6 antibody.
      Ten percent of the precipitated proteins (lane 1: mouse IgG, lane 2: Gemin6) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Gemin6 antibody (1:5000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232)
      Arrow indicates Gemin6 (Please see figures).

      Western Blot Analysis:
      Representative lot data.
      HeLa cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-Gemin6 (0.1 µg/mL).
      Proteins were visualized using a Goat Anti-Mouse IgG conjugated to HRP and chemiluminescence detection system.
      Arrow indicates Gemin6 (~16 kDa) (Please see figures).


      Immunoprecipitation:
      A representative lot was used by an independent laboratory for IP (Battle, D.J., et al. (2007). The Journal of Biological Chemistry. 282:27953-27959).
      Biological Information
      ImmunogenRecombinant protein corresponding to human Gemin6.
      HostMouse
      SpecificityRecognizes Gemin6.
      IsotypeIgG1κ
      Species Reactivity
      • Human
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryGEMIN6 is part of a large macromolecular complex, localized to both the cytoplasm and the nucleus, that plays a role in the cytoplasmic assembly of small nuclear ribonucleoproteins (snRNPs). Other members of this complex include SMN (MIM 600354), GEMIN2 (SIP1; MIM 602595), GEMIN3 (DDX20; MIM 606168), GEMIN4 (MIM 606969), and GEMIN5 (MIM 607005).[supplied by OMIM].
      Gene Symbol
      • GEMIN6
      • SIP2
      Purification MethodProtein G Purified
      UniProt Number
      UniProt SummaryFUNCTION: The SMN complex plays an essential role in spliceosomal snRNP assembly in the cytoplasm and is required for pre-mRNA splicing in the nucleus.

      SUBUNIT STRUCTURE: Part of the core SMN complex that contains SMN1, SIP1/GEMIN2, DDX20/GEMIN3, GEMIN4, GEMIN5, GEMIN6, GEMIN7, GEMIN8 and STRAP/UNRIP. Interacts directly with GEMIN7, SNRPB, SNRPD2, SNRPD3 and SNRPE.

      SUBCELLULAR LOCATION: Nucleus › nucleoplasm. Nucleus › gem. Cytoplasm. Note: Found both in the nucleoplasm and in nuclear bodies called gems (Gemini of Cajal bodies) that are often in proximity to Cajal (coiled) bodies. Also found in the cytoplasm.

      SEQUENCE CAUTION: The sequence BAB15660.1 differs from that shown. Reason: Frameshift at position 142.
      Molecular Weight ~16 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRNA 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-Gemin6 antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
      Successful immunoprecipitation of Gemin6-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.
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsStable 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.
      Packaging Information
      Material Size10 assays
      Material Package10 assays per set. Recommended use: ~5 μg of antibody per RIP (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

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      Categories

      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies