03-101 | RIPAb+ PABPC1 - RIP Validated Antibody and Primer Set

03-101
10 assays  10 assays per set. Recommended use: ~5 μg antibody per RIP (dependent upon biological context).
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey Applications
      H, Rb, XnRIP, IP, WB
      Description
      Catalogue Number03-101
      Brand Family Upstate
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+ PABPC1 - RIP Validated Antibody and Primer Set
      OverviewRIPAb+ antibodies are evaluated using the RNA Binding Protein Immunoprecipitation (RIP) assay. Each RIPAb+ antibody set includes quantitative RT-PCR control primers (RIP Primers) to biologically validate your IP results by successfully coprecipitating a specific RNA target (where such a specific target is known). The qPCR protocol and primer sequences are provided, allowing researchers to validate RIP protocols when using the antibody in their experimental context. Each set also includes a negative control antibody to ensure specificity of the RIP reaction.
      The RIPAb+ PABPC1 set includes the PABPC1 antibody, a negative control antibody (purified mouse IgG), and positive qPCR primers which amplify a 100 bp fragment of the human ACTB cDNA. The PABPC1 and negative control antibodies are supplied in a scalable "per RIP" reaction size and can be used to functionally validate the precipitation of PABPC1-associated RNAs.
      References
      Product Information
      FormatPurified
      Control
      • Included negative control mouse IgG antibody and control primers specific for actin B.
      PresentationAnti-PABPC1, clone 10E10 (mouse monoclonal IgG)
      One vial 50 μg of protein A purified mouse IgG2b in 50 μL of PBS containing 0.1% sodium azide. 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, ACTB
      One vial containing 75 μL of 5 μM of each primer specific for actin B (ACTB). Store at -20°C.
      FOR: TTG TTA CAG GAA GTC CCT TGC C
      REV: ATG CTA TCA CCT CCC CTG TGT G
      Applications
      ApplicationThis RIPAb+ PABPC1 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
      Key Applications
      • RNA Binding Protein Immunoprecipitation (RIP)
      • Immunoprecipitation
      • Western Blotting
      Application NotesImmunoprecipitation from RIP lysate:
      RIP lysate from HeLa cells (2 X 106 cell equivalents per IP) was subjected to immunoprecipitation using 0.5 μg of either a normal mouse IgG or Anti-PABPC1 antibody. Precipitated proteins were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-PABPC1 (1.0 μg/mL). Proteins were visualized using One-Step™ IP-Western kit (GenScript Cat. # L00232) (Please see figures).

      Western Blot Analysis:
      0.5-2 μg/mL of a previous lot detected PABPC1 in RIPA lysates from HeLa and Jurkat cells (Please see figures).
      Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-PABPC1 (0.5 μg/mL). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
      Biological Information
      ImmunogenThe PABPC1 monoclonal antibody is made against a fusion protein corresponding to full-length of human cytoplasmic poly (A)-binding protein (PABPC1).
      Clone10E10
      HostMouse
      SpecificityPABPC1
      IsotypeIgG2b
      Species Reactivity
      • Human
      • Rabbit
      • Xenopus
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThe poly(A)-binding protein (PABP), which is found complexed to the 3-prime poly(A) tail of eukaryotic mRNA, is required for poly(A) shortening and translation initiation. In humans, the PABPs comprise a small nuclear isoform and a conserved gene family that displays at least 3 functional proteins: PABP1 (PABPC1), inducible PABP (iPABP, or PABPC4; MIM 603407), and PABP3 (PABPC3; MIM 604680). In addition, there are at least 4 pseudogenes, PABPCP1 to PABPCP4.[supplied by OMIM]
      Gene Symbol
      • PABPC1
      • PABP1
      • PABPC2
      • PABPL1
      • PAB1
      • Poly
      • PABP
      Purification MethodProtein A Purfied
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P11940 # Binds the poly(A) tail of mRNA. May be involved in cytoplasmic regulatory processes of mRNA metabolism such as pre- mRNA splicing. Its function in translational initiation regulation can either be enhanced by PAIP1 or repressed by PAIP2. Can probably bind to cytoplasmic RNA sequences other than poly(A) in vivo. May be involved in translationally coupled mRNA turnover. Implicated with other RNA-binding proteins in the cytoplasmic deadenylation/translational and decay interplay of the FOS mRNA mediated by the major coding-region determinant of instability (mCRD) domain.
      SIZE: 636 amino acids; 70671 Da
      SUBUNIT: Component of a multi subunit autoregulatory ribonucleoprotein complex (ARC), at least composed of IGF2BP1, PABPC1 and CSDE1. Interacts with IGF2BP1. Part of a complex associated with the FOS mCRD domain and consisting of HNRPD, SYNCRIP, PAIP1 and CSDE1/UNR. Interacts with the PABPC1- interacting motif-1 (PAM1) and -2 (PAM2) of PAIP1 and PAIP2. Interacts with PAIP1 with a 1:1 stoichiometry and with PAIP2 with a 1:2 stoichiometry. The interaction with CSDE1 is direct and RNA- independent. Found in a mRNP complex with YBX2. 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, HNRPA1, HNRPA2B1, HNRPA3, HNRPC, HNRPF, HNRPH1, HNRPK, HNRPM, HNRPR, HNRPU, 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 (By similarity).
      SUBCELLULAR LOCATION: Cytoplasm. Nucleus. Note=Shuttles between the cytoplasm and the nucleus.
      TISSUE SPECIFICITY: Ubiquitous.
      DOMAIN: "SwissProt: P11940 The RNA-binding domains RRM1 and RRM2 and the C-terminus (last 138 amino acids) regions interact with the PABPC1- interacting motif-1 (PAM1) and -2 (PAM2) of PAIP1, respectively. & The RNA-binding domains RRM2 and RRM3 and the C-terminus (last 138 amino acids) regions interact with the PABPC1- interacting motif-1 (PAM1) and -2 (PAM2) of PAIP2, respectively.
      PTM: Methylated by CARM1. & Arg-493 is dimethylated, probably to asymmetric dimethylarginine.
      SIMILARITY: Contains 1 PABC domain. & Contains 4 RRM (RNA recognition motif) domains.
      Molecular Weight73 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 107 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or the anti-PABPC1 antibody and the Magna RIP Kit (Cat. # 17-700).
      Successful immunoprecipitation of PABPC1-associated RNA was verified by qRT-PCR using RIP Primers, ACTB (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 first thaw, 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.
      Packaging Information
      Material Size10 assays
      Material Package10 assays per set. Recommended use: ~5 μg antibody per RIP (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Certificates of Analysis

      TitleLot Number
      RIPAb+ PABPC1, clone 10E10 3082347
      RIPAb+ PABPC1, clone 10E10 - 2397085 2397085
      RIPAb+ PABPC1, clone 10E10 - 2537174 2537174
      RIPAb+ PABPC1, clone 10E10 - JH1752521 JH1752521
      RIPAb+ PABPC1, clone 10E10 - NG1627193 NG1627193
      RIPAb+ PABPC1, clone 10E10 -2642151 2642151
      RIPAb+ PABPC1, clone 10E10 -2654205 2654205
      RIPAb+ PABPC1, clone 10E10 -2743633 2743633
      RIPAb+ PABPC1, clone 10E10 -2757906 2757906
      RIPAb+ PABPC1, clone 10E10 Monoclonal Antibody/Primer Set 3085267

      Brochure

      Title
      RNA-Binding Protein Immunoprecipitation

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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