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NRD1 Antikörper (Middle Region)

NRD1 Reaktivität: Human, Pferd, Schwein, Rind (Kuh) WB Wirt: Kaninchen Polyclonal unconjugated
Produktnummer ABIN2787813
  • Target Alle NRD1 Antikörper anzeigen
    NRD1 (Nardilysin (N-Arginine Dibasic Convertase) (NRD1))
    Bindungsspezifität
    • 7
    • 6
    • 6
    • 6
    • 5
    • 3
    • 3
    • 1
    • 1
    • 1
    Middle Region
    Reaktivität
    Human, Pferd, Schwein, Rind (Kuh)
    Wirt
    • 26
    • 1
    Kaninchen
    Klonalität
    • 29
    Polyklonal
    Konjugat
    • 15
    • 4
    • 3
    • 3
    • 2
    • 2
    Dieser NRD1 Antikörper ist unkonjugiert
    Applikation
    • 23
    • 23
    • 4
    • 1
    • 1
    Western Blotting (WB)
    Sequenz
    GSKMLSVHVV GYGKYELEED GTPSSEDSNS SCEVMQLTYL PTSPLLADCI
    Homologie
    Cow: 93%, Horse: 86%, Human: 100%, Pig: 93%
    Produktmerkmale
    This is a rabbit polyclonal antibody against NRD1. It was validated on Western Blot using a cell lysate as a positive control.
    Aufreinigung
    Affinity Purified
    Immunogen
    The immunogen is a synthetic peptide directed towards the middle region of human NRD1
  • Applikationshinweise
    Optimal working dilutions should be determined experimentally by the investigator.
    Kommentare

    Antigen size: 1151 AA

    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Validierung #103724 (Immunohistochemistry)
    'Independent Validation' Siegel
    by
    AG Deeg, Lehrstuhl für Physiologie, Veterinärwissenschaftliches Department, Tierärztliche Fakultät, Ludwig-Maximilians-Universität München
    No.
    #103724
    Datum
    19.07.2019
    Antigen
    NRD1
    Chargennummer
    QC27244-100203
    Validierte Anwendung
    Immunohistochemistry
    Positivkontrolle

    retina from horses suffering from equine recurrent uveitis (ERU)

    Negativkontrolle

    retina from non-ERU infected horses

    isolated equine retinal Müller glial cells (eqMCs)

    Bewertung

    Passed. The NRD1 antibody ABIN2787813 specifically labels the targeted antigen in retinae from horses suffering from ERU and in isolated equine retinal Müller glial cells.

    'Independent Validation' Siegel
    Validierungsbilder
    Protokoll
    Primärantikörper
    ABIN5074774
    Sekundärantikörper
    goat anti-rabbit IgG (H+L) AF568-conjugated antibody (Invitrogen, A11036, lot 1832035)
    Full Protocol
    • IHC on fixed equine retina:
      • Dissect tissue and fix in 4% buffered formalin for 12h at 4°C.
      • Dehydrate and paraffinize the tissue through a graded alcohole, xylene and paraffine series.
      • Embed the tissue in Richard Allen Scientific Paraffine Type 9 (Thermofisher, 8337).
      • Cut fixed tissue with a microtome into 8µm-thick sections.
      • Deparaffinization and rehydration through graded xylene and graded alcohol series:
      • Boil the sections in a 99°C hot waterbath in 0.1M Na2-EDTA buffer pH8.0 for 15min.
      • Boil the sections in a 99°C hot waterbath in citrate buffer pH6 (Dako, 2031) for 15min.
      • Rinse sections with PBS pH7.4.
    • IHC on isolated eqMCs:
      • Prepare eqMCs from the eyes of a 19 year old mare.
      • Cultivate cells in DMEM (PAN-Biotech, P04-04510, lot 2650219) supplemented with 10% v/V FBS (Biochrom, S0615, lot 0879F) and 1% Penicillin/Streptomycin (PAN-Biotech, P06-07100, lot 7180417) for 18d at 37°C at 5% CO2.
      • Seed 1x105 cells (first passage) in 1ml medium on adhesion microscope slides (Langenbrinck 03-0060, lot 021218).
      • Incubate cells for 2d at 37°C at 5% CO2.
      • Remove medium and wash cells with PBS pH7.4.
      • Fix cells for 10min with ice-cold acetone.
      • Rehydrate cells with TBST (0.01M Tris, 0.15M NaCl, 0.1% v/v Tween 20) pH7.3.
    • Block samples in 5% goat serum in TBST pH7.3 containing 1% w/v BSA (BSA-TBST).
    • Incubate samples with primary rabbit anti-NRD1 antibody (antibodies-online, ABIN2787813, lot QC27244-100203) diluted 1:100 in BSA-TBST ON at 4°C.
    • Wash samples with BSA-TBST.
    • Incubate samples with secondary goat anti-rabbit IgG (H+L) AF568-conjugated antibody (Invitrogen, A11036, lot 1832035) diluted 1:500 in BSA-TBST for 30min at RT.
    • Wash samples with BSA-TBST.
    • Incubate samples with primary mouse anti-Vimentin antibody (Sigma-Aldrich, V6630) diluted 1:400 in BSA-TBST ON at 4°C.
    • Wash samples with BSA-TBST.
    • Incubate samples with secondary goat anti-mouse IgG (H+L) AF488-conjugated antibody (Invitrogen, A11029, lot 1874804) diluted 1:500 in BSA-TBST containing 0.1% DAPI for 30min at RT.
    • Wash samples with BSA-TBST.
    • Mount samples in Fluoromount W mounting medium (SERVA, 21634, lot 181008).
    • Acquire images with Leica DMi8 fluorescence microscope equipped with Filter Cubes for DAPI, FITC, TXR and Y5 at 200x (prepared retina) and 400x (eqMCs). Parameters for image acquisition (exposure: FITC 220ms, TXR 253ms, DAPI 195ms) were maintained unchanged for all images.
    Anmerkungen

    ABIN2787813 was also tested in western blot on retinal Müller glial cell lysate from healthy horses or horses suffering from ERU and in equine and porcine total retinal lysates. However, the antibody did not detect a protein of the expected MW of 132kDa in any of these samples whereas numerous extraneous protein bands were detected.

  • Format
    Liquid
    Konzentration
    Lot specific
    Buffer
    Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.
    Konservierungsmittel
    Sodium azide
    Vorsichtsmaßnahmen
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Handhabung
    Avoid repeated freeze-thaw cycles.
    Lagerung
    -20 °C
    Informationen zur Lagerung
    For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.
  • Target
    NRD1 (Nardilysin (N-Arginine Dibasic Convertase) (NRD1))
    Andere Bezeichnung
    NRD1 (NRD1 Produkte)
    Synonyme
    hNRD1 antikoerper, hNRD2 antikoerper, NRDC antikoerper, 2600011I06Rik antikoerper, AI875733 antikoerper, NRD-C antikoerper, NRD1 antikoerper, si:dkey-171o17.4 antikoerper, DKFZp459N143 antikoerper, nardilysin convertase antikoerper, nardilysin, N-arginine dibasic convertase, NRD convertase 1 antikoerper, nardilysin antikoerper, nardilysin b (N-arginine dibasic convertase) antikoerper, NRDC antikoerper, Nrdc antikoerper, Nrd1 antikoerper, LOC552055 antikoerper, nrd1b antikoerper, CpipJ_CPIJ002524 antikoerper, Tsp_03755 antikoerper
    Hintergrund
    NRD1 cleaves peptide substrates on the N-terminus of arginine residues in dibasic pairs.
    Alias Symbols: hNRD1, hNRD2
    Protein Interaction Partner: FBXW11, UBC, rev, SETSIP, PUS1, P3H1, ELAC2, XPO5, NSUN2, PDIA6, YARS, DNAJC7, TSN, PPP3CA, PPP2R2A, PPP2CA, PFDN5, PFDN1, PPP1R12A, MSH2, PNKP, HSPB1, MOB2, MOB1A, STK3, NRD1, HIST2H3C, UACA, COBLL1, NCOR2, NCOR1, HDAC3, IQGAP1, MAD1L1, TBL1X, UIMC1, BTR
    Protein Size: 1151
    Molekulargewicht
    132 kDa
    Gen-ID
    4898
    NCBI Accession
    NM_001101662, NP_001095132
    UniProt
    O43847
    Pathways
    Skeletal Muscle Fiber Development
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