03-242 RIPAb+™ PUM1 Antibody

03-242
10 assays  10 assays per set. Recommended use: ~2.5 µg (7.2 µL) of antibody per RIP (dependent upon biological context).
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey Applications
      H, M, R, B, Rhesus Macaque, Chp, Pm ICC, WB, RIP
      Description
      Catalogue Number03-242
      Trade Name
      • RIPAb+
      • Upstate
      DescriptionRIPAb+™ PUM1 Antibody
      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.
      PUM1 has been shown to bind an mRNA sequence known as Nanos response element. It is found in heart, brain, muscle, kidney, stomach and intestine. PUM proteins play an active role in the sustainability of mitotic stem cell proliferation and the regulation of cell development and differentiation.
      Alternate Names
      • pumilio (Drosophila) homolog 1
      • pumilio 1
      • pumilio homolog 1 (Drosophila)
      Background InformationWithin the mammalian genome are two genes belonging to a family of housekeeping genes known as PUF. These genes are known as PUM1 and PUM2. PUF proteins are characterized by having a C-terminal RNA-binding domain called the Pumilio homolog domain, which affects the stability of mRNA and translation by binding the 3’-UTR of mRNA targets. PUM plays an active role in the sustainability of mitotic stem cell proliferation and the regulation of cell development and differentiation. Very little has been discovered about the specific role for PUM1. However it has been shown to bind an mRNA sequence known as Nanos response element and to interact with RNA. PUM1 is found in heart, brain, muscle, kidney, stomach and intestine. It is not expressed in putamen, hippocampus, medulla oblangata, corpus collasum, cerebellum and caudate nucleus.
      References
      Product Information
      FormatAffinity Purified
      Control
      • Includes normal rabbit IgG and primers specific for human p27 mRNA.
      PresentationComponents:
      Anti-PUM1 (Rabbit Polyclonal). One vial containing 25 µg of affinity purified polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl,with 0.05% sodium azide before the addition of 30% glycerol. Store at -20°C.
      Concentration: 0.35 mg/mL
      Normal Rabbit IgG. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide. Store at -20°C.
      RIP Primers, p27 mRNA. One vial containing 75 μL of each primer (5 μM) specific for a region of the human p27 (Kip1/CDKN1B) mRNA. Store at -20°C.
      FOR: CTG CAA CCG ACG ATT CTT CTA CT
      REV: GGG CGT CTG CTC CAC AGA
      Applications
      ApplicationRIPAb+ PUM1 Antibody detects level of RIPAb+ PUM1 & has been published & validated for use in ICC, Western Blotting, RNA Binding Protein Immunoprecipitation (RIP).
      Key Applications
      • Immunocytochemistry
      • Western Blotting
      • RNA Binding Protein Immunoprecipitation (RIP)
      Application NotesWestern Blot Analysis:
      Representative lot data.
      H9 human stem cell lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-Pumilio-1 (PUM1) (1:5,000 dilution).
      Proteins were visualized using a Donkey Anti-Rabbit IgG conjugated to HRP and chemiluminescence detection system.
      Arrow indicates Pumilio-1 (~120 kDa).
      Immunocytochemistry Analysis:
      Representative lot data.
      Confocal fluorescent analysis of mouse and human embryonic stem cells using Anti-Pumilio-1 (PUM1) (Red). Actin filaments have been labeled with Alexa Fluor® 488 dye - Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
      Biological Information
      ImmunogenKLH-conjugated linear peptide corresponding to the N-terminus of human Pumilio-1.
      EpitopeN-terminus
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostRabbit
      SpecificityThis antibody recognizes the N-terminus of Pumilio-1 (PUM1).
      Species Reactivity
      • Human
      • Mouse
      • Rat
      • Bovine
      • Rhesus Macaque
      • Chimpanzee
      • Primate
      Species Reactivity NoteDemonstrated to react with human, mouse, and rat.
      Predicted to react with bovine, rhesus monkey, chimpanzee, and primate based on 100% sequence homology.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a member of the PUF family, evolutionarily conserved RNA-binding proteins related to the Pumilio proteins of Drosophila and the fem-3 mRNA binding factor proteins of C. elegans. The encoded protein contains a sequence-specific RNA binding domain comprised of eight repeats and N- and C-terminal flanking regions, and serves as a translational regulator of specific mRNAs by binding to their 3' untranslated regions. The evolutionarily conserved function of the encoded protein in invertebrates and lower vertebrates suggests that the human protein may be involved in translational regulation of embryogenesis, and cell development and differentiation. Alternatively spliced transcript variants encoding different isoforms have been described. [provided by RefSeq].
      Gene Symbol
      • PUM1
      • HSPUM
      • HsPUM
      • PUMH
      • PUMH1
      • PUML1
      • Pumilio-1
      Purification MethodAffinity Purfied
      UniProt Number
      UniProt SummaryFUNCTION: Sequence-specific RNA-binding protein that regulates translation and mRNA stability by binding the 3'-UTR of mRNA targets. May be required to support proliferation and self-renewal of stem cells By similarity.

      SUBUNIT STRUCTURE: Binds in a sequence-specific manner to the 3'-UTR of some mRNAs.

      SUBCELLULAR LOCATION: Cytoplasm Probable.

      TISSUE SPECIFICITY: Expressed in brain, heart, kidney, muscle, intestine and stomach. Not expressed in cerebellum, corpus callosum, caudate nucleus, hippocampus, medulla oblongata and putamen. Expressed in all fetal tissues tested.

      INDUCTION: Strongly down-regulated in keratinocytes upon ultraviolet B radiation.

      DOMAIN: The pumilio repeats mediate the association with RNA by packing together to form a right-handed superhelix that approximates a half donut. The number as well as the specific sequence of the repeats determine the specificity for target mRNAs.

      SEQUENCE SIMILARITIES: Contains 1 PUM-HD domain.

      Contains 8 pumilio repeats.
      Molecular Weight~120 kDa.
      UniProtKB/Swiss-Prot Q14671 (PUM1_HUMAN) entry describes 2 isoforms produced by alternative splicing: Isoform 1 at ~126 kDa and Isoform 2 at ~124 kDa.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRNA Binding Protein Immunoprecipitation:
      Representative lot data.
      RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 2.5 µg of either a normal Rabbit IgG or 2.5 µg Anti-PUM1 antibodyand the Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of PUM1-associated RNA was verified by qPCR using RIP Primers p27 mRNA, (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 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.
      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: ~2.5 µg (7.2 µL) of antibody per RIP (dependent upon biological context).
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      RIPAb+ PUM1 - Q1975311Q1975311
      RIPAb+™ PUM12463677
      RIPAb+™ PUM1 -27525122752512
      RIPAb+™ PUM1 Polyclonal Antibody/Primer Set2986330

      References

      Reference overviewPub Med ID
      Ribonomic analysis of human Pum1 reveals cis-trans conservation across species despite evolution of diverse mRNA target sets.
      Morris, Adam R, et al.
      Mol. Cell. Biol., 28: 4093-103 (2008)  2008

      Show Abstract
      18411299 18411299
      Dendritic localization of the translational repressor Pumilio 2 and its contribution to dendritic stress granules.
      Vessey, John P, et al.
      J. Neurosci., 26: 6496-508 (2006)  2006

      Show Abstract
      16775137 16775137
      Evidence for distinct pathomechanisms in B-cell chronic lymphocytic leukemia and mantle cell lymphoma by quantitative expression analysis of cell cycle and apoptosis-associated genes.
      Korz, Christian, et al.
      Blood, 99: 4554-61 (2002)  2002

      Show Abstract
      12036888 12036888
      The Drosophila pumilio gene encodes two functional protein isoforms that play multiple roles in germline development, gonadogenesis, oogenesis and embryogenesis.
      Parisi, M and Lin, H
      Genetics, 153: 235-50 (1999)  1999

      Show Abstract
      10471709 10471709

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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