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03-181 RIPAb+™ hnRNPA1 (M9 Region) - RIP Validated Antibody and Primer Set

03-181
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 Specifications Table

      Species ReactivityKey Applications
      HRIP, WB, IP
      Description
      Catalogue Number03-181
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+™ hnRNPA1 (M9 Region) - 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.
      The hnRNPs are RNA binding proteins that complex with heterogeneous nuclear RNA (hnRNA) and associate with pre-mRNAs in the nucleus. These complexes are associated with pre-mRNA processing and other aspects of mRNA metabolism and transport. The hnRNP A1 protein is involved in the packaging of pre-mRNA into hnRNP particles, transport of poly A+ mRNA from the nucleus to the cytoplasm, and may have a role in splice site selection.
      Alternate Names
      • Helix-destabilizing protein
      • Single-strand RNA-binding protein
      • heterogeneous nuclear ribonucleoprotein A1
      • heterogeneous nuclear ribonucleoprotein A1B protein
      • heterogeneous nuclear ribonucleoprotein B2 protein
      • heterogeneous nuclear ribonucleoprotein core protein A1
      • hnRNP core protein A1
      • nuclear ribonucleoprotein particle A1 protein
      • single-strand DNA-binding protein UP1
      Background InformationThe hnRNPs are RNA binding proteins that complex with heterogeneous nuclear RNA (hnRNA) and associate with pre-mRNAs in the nucleus. These complexes are associated with pre-mRNA processing and other aspects of mRNA metabolism and transport. While all hnRNPs are present in the nucleus, data suggests they shuttle between the nucleus and the cytoplasm. The hnRNP proteins have distinct nucleic acid binding properties. The protein encoded by hnRNP A1 has two repeats of quasi-RRM domains that bind to RNAs. This abundant core protein, along with other hnRNP proteins, is exported from the nucleus, probably bound to mRNA, and is immediately re-imported. The hnRNP A1 protein is involved in the packaging of pre-mRNA into hnRNP particles, transport of poly A+ mRNA from the nucleus to the cytoplasm, and may have a role in splice site selection.
      References
      Product Information
      FormatPurified
      Control
      • Includes negative control mouse IgG antibody and control primers specific for the mRNA of human E2F3.
      PresentationAnti-hnRNPA1 (M9 Region) (Mouse Monoclonal). One vial containing 50 μg of purified antibody in 75 μL of 0.1 M Tris-Glycine (pH7.4), 150 mM NaCl with 0.05% sodium azide and 30% glycerol. 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, E2F3, . One vial containing 75 μL of 5 μM of each primer specific for human E2F3 mRNA.
      Store at -20°C.
      FOR: TGG TAC CAT TGA GTT GCT GCT ATT
      REV: AGC TCA TGT GTT GCC CTT TAT ACA
      Quality LevelMQ100
      Applications
      ApplicationThis RIPAb+ hnRNPA1 (M9 Region) -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • RNA Binding Protein Immunoprecipitation (RIP)
      • Western Blotting
      • 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 5 µg of either a normal mouse IgG, (Cat. #CS200621), or 5 µg of Anti-hnRNPA1 (M9 Region) antibody (Cat. # CS204410). Ten percent of the precipitated proteins (lane 1: mouse IgG, lane 3: hnRNPA1) and hela whole cell lysate (lane 2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-hnRNPA1 (M9 Region) antibody (Cat. # CS204410, 1:1000). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232)
      Arrow indicates hnRNPA1 (~38 kDa) (Figure 2).
      Western Blot Analysis:
      Representative lot data.
      Hela nuclear extract was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-hnRNP A1, clone 9H10 (0.01 µg/mL).
      Proteins were visualized using a Goat Anti-Mouse IgG conjugated to HRP and chemiluminescence detection system.
      Arrow indicates hnRNPA1 (~38 kDa) (Figure 3).
      Biological Information
      ImmunogenRecombinant protein corresponding to human hnRNP A1.
      EpitopeM9 Region
      Clone9H10
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityThis antibody recognizes hnRNP A1.
      IsotypeIgG2bκ
      Species Reactivity
      • Human
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene belongs to the A/B subfamily of ubiquitously expressed heterogeneous nuclear ribonucleoproteins (hnRNPs). The hnRNPs are RNA binding proteins and they complex with heterogeneous nuclear RNA (hnRNA). These proteins are associated with pre-mRNAs in the nucleus and appear to influence pre-mRNA processing and other aspects of mRNA metabolism and transport. While all of the hnRNPs are present in the nucleus, some seem to shuttle between the nucleus and the cytoplasm. The hnRNP proteins have distinct nucleic acid binding properties. The protein encoded by this gene has two repeats of quasi-RRM domains that bind to RNAs. It is one of the most abundant core proteins of hnRNP complexes and it is localized to the nucleoplasm. This protein, along with other hnRNP proteins, is exported from the nucleus, probably bound to mRNA, and is immediately re-imported. Its M9 domain acts as both a nuclear localization and nuclear export signal. The encoded protein is involved in the packaging of pre-mRNA into hnRNP particles, transport of poly A+ mRNA from the nucleus to the cytoplasm, and may modulate splice site selection. It is also thought have a primary role in the formation of specific myometrial protein species in parturition. Multiple alternatively spliced transcript variants have been found for this gene but only two transcripts are fully described. These variants have multiple alternative transcription initiation sites and multiple polyA sites. [provided by RefSeq].
      Gene Symbol
      • HNRNPA1
      • HNRPA1
      • hnRNPA1
      Purification MethodProtein G Purified
      UniProt Number
      UniProt SummaryFUNCTION: Involved in the packaging of pre-mRNA into hnRNP particles, transport of poly(A) mRNA from the nucleus to the cytoplasm and may modulate splice site selection. May play a role in HCV RNA replication.

      SUBUNIT STRUCTURE: Identified in the spliceosome C complex, at least composed of AQR, ASCC3L1, C19orf29, CDC40, CDC5L, CRNKL1, DDX23, DDX41, DDX48, DDX5, DGCR14, DHX35, DHX38, DHX8, EFTUD2, FRG1, GPATC1, HNRNPA1, HNRNPA2B1, HNRPA3, HNRNPC, HNRPF, HNRPH1, HNRPK, HNRPM, HNRNPR, HNRNPU, KIAA1160, KIAA1604, LSM2, LSM3, MAGOH, MORG1, PABPC1, PLRG1, PNN, PPIE, PPIL1, PPIL3, PPWD1, PRPF19, PRPF4B, PRPF6, PRPF8, RALY, RBM22, RBM8A, RBMX, SART1, SF3A1, SF3A2, SF3A3, SF3B1, SF3B2, SF3B3, SFRS1, SKIV2L2, SNRPA1, SNRPB, SNRPB2, SNRPD1, SNRPD2, SNRPD3, SNRPE, SNRPF, SNRPG, SNW1, SRRM1, SRRM2, SYF2, SYNCRIP, TFIP11, THOC4, U2AF1, WDR57, XAB2 and ZCCHC8. Identified in a mRNP granule complex, at least composed of ACTB, ACTN4, DHX9, ERG, HNRNPA1, HNRNPA2B1, HNRNPAB, HNRNPD, HNRNPL, HNRNPR, HNRNPU, HSPA1, HSPA8, IGF2BP1, ILF2, ILF3, NCBP1, NCL, PABPC1, PABPC4, PABPN1, RPLP0, RPS3, RPS3A, RPS4X, RPS8, RPS9, SYNCRIP, TROVE2, YBX1 and untranslated mRNAs. Interacts with SEPT6. Interacts with HCV NS5B and with the 5'-UTR and 3'-UTR of HCV RNA.

      SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note: Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Shuttles continuously between the nucleus and the cytoplasm along with mRNA. Component of ribonucleosomes. In the course of viral infection, colocalizes with HCV NS5B at speckles in the cytoplasm in a HCV-replication dependent manner.

      PTM: Arg-194, Arg-206 and Arg-225 are dimethylated, probably to asymmetric dimethylarginine.

      SEQUENCE SIMILARITIES: Contains 2 RRM (RNA recognition motif) domains.
      Molecular Weight~ 38 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 5 µg of either a normal mouse IgG , or 5 µg Anti-hnRNPA1 (M9 Region) antibody and the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700).
      Successful immunoprecipitation of hnRNPA1-associated RNA was verified by qPCR using RIP Primers E2F3, (Figure 1).
      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 > Antibodies and Assays > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies
      Life Science Research > Antibodies and Assays > Primary Antibodies
      Life Science Research > Protein Detection and Quantification > Immunoassays > Immunoprecipitation (IP) > RNA-Binding Protein Immunoprecipitation (RIP) > RIP Validated Antibodies