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Plakophilin 2 Antikörper (N-Term)

PKP2 Reaktivität: Human, Ratte, Maus, Hund, Rind (Kuh), Pferd WB, IHC Wirt: Kaninchen Polyclonal unconjugated
Produktnummer ABIN2786784
  • Target Alle Plakophilin 2 (PKP2) Antikörper anzeigen
    Plakophilin 2 (PKP2)
    Bindungsspezifität
    • 15
    • 8
    • 7
    • 5
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    N-Term
    Reaktivität
    • 50
    • 17
    • 17
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Human, Ratte, Maus, Hund, Rind (Kuh), Pferd
    Wirt
    • 42
    • 5
    • 2
    • 1
    Kaninchen
    Klonalität
    • 45
    • 5
    Polyklonal
    Konjugat
    • 28
    • 4
    • 4
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Dieser Plakophilin 2 Antikörper ist unkonjugiert
    Applikation
    • 37
    • 23
    • 22
    • 6
    • 5
    • 5
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunohistochemistry (IHC)
    Sequenz
    HPLRRLEISP DSSPERAHYT HSDYQYSQRS QAGHTLHHQE SRRAALLVPP
    Homologie
    Cow: 77%, Dog: 79%, Horse: 86%, Human: 100%, Mouse: 86%, Rat: 77%
    Produktmerkmale
    This is a rabbit polyclonal antibody against PKP2. 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 N terminal region of human PKP2
    Top Product
    Discover our top product PKP2 Primärantikörper
  • Applikationshinweise
    Optimal working dilutions should be determined experimentally by the investigator.
    Kommentare

    Antigen size: 837 AA

    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Validierung #103654 (Western Blotting)
    'Independent Validation' Siegel
    by
    Gencardio (IDIBGI)
    No.
    #103654
    Datum
    13.03.2019
    Antigen
    PKP2
    Chargennummer
    QC27639-43357
    Validierte Anwendung
    Western Blotting
    Positivkontrolle

    HL-1 cell extracts (from AT-1 mouse atrial cardiomyocyte tumor lineage)

    Extracts from HEK293 cells transfected with human PKP2-FLAG expressing plasmid

    Extract of mouse heart tissue (endogenous PKP2 expected molecular weight 88KDa)

    Extract of human heart tissue (endogenous PKP2 expected molecular weight 91KDa)

    Negativkontrolle

    Extracts from untransfected HEK293 cells

    Bewertung

    ABIN2786784 specifically recognizes PKP2 in cell extracts from HEK-293 overexpressing human PKP2 protein and endogenous PKP2 in human heart tissue but not endogenous mouse PKP2 neither in HL-1 cells nor mouse tissue.

    'Independent Validation' Siegel
    Validierungsbilder
    Protokoll
    Primärantikörper
    ABIN2786784
    Sekundärantikörper
    mouse anti-FLAG-tag antibody (Sigma, monoclonal anti-flag, F316, lot SLBT7654)
    Full Protocol
    • Grow HL-1 cells in Claycomb medium (Sigma, 51800C, lot SLBX8700) supplemented with 10% FBS (Gibco, 10270-106, lot 42G5178K), 1% Penicillin-Streptomycin (Sigma, P4333, lot 048M4874V) and 1% Glutamax (Gibco, 35050-061, 2037045), at 37°C and 5% CO2 in 2ml on a 6 well plate to 95% confluency.
    • Grow HEK293 cells in DMEM/High Modified medium (GE Life Science, SH30285.01, lot AC14565263) supplemented with 10% FBS, 1% Penicillin-Streptomycin and 1% Glutamax (Gibco, 35050-061, 2037045), at 37°C and 5% CO2 in 2ml on a 6 well plate to 95% confluency.
    • Transfect HEK293 cells with PKP2a plasmid (p915 PKP2a-pFLAG-cmv5a; provided by Dr. Green KJ) using Lipofectamin 2000 (Invitrogen, 11668-019, lot 2022713) following the manufacturer´s instructions.
    • Lyse approximately 1.2x106 cells per well in 200µl 1% SDS (Sigma, 75746, lot STBG4187V).
    • Homogenate whole heart of transgenic and wt heart mouse tissue with M-Per reagent (Thermo, 78501, lot NC168900) and mechanical disgregation using a Dounce homogenizer.
    • Incubate for the lysates from 15min at 95°C.
    • Vortex for 15min.
    • Determine total protein content of the lysates using Pierce BCA Protein Assay Kit (Thermo Scientific, 23227, lot TC263613).
    • Denature 15µg (HEK293 cell lysates), 106µg (HL-1 cell lysates), and 100µg (mouse heart tissue lysates) of total protein for 5min at 95°C in 45µl using 6x Laemmli SDS sample buffer and subsequently separate them on a 8% acrylamide gel in an electrophoresis chamber for 30min at 80V and 1h at 120V.
    • Transfer proteins onto Amersham Hybond P 0.45 PVDF blotting membrane (GE Life Science, 10600023, lot A16954274) for 2h at 80V.
    • Block the membrane with 5% NFM in PBST for 1h at RT.
    • Incubation with primary
      • rabbit anti-Plakophilin 2 (PKP2) (N-Term) antibody (antibodies-online, ABIN2786784, lot QC27639-43357) diluted 1:800 in 5% NFM in PBS ON at 4°C or
      • mouse anti-FLAG-tag antibody (Sigma, monoclonal anti-flag, F316, lot SLBT7654) diluted 1:x in 5% NFM in PBS ON at 4°C.
    • Wash membrane 2x for 5’ with PBST and 2x for 5min with PBS.
    • Block the membrane with 5% NFM in PBST for 1h at RT.
    • Incubation with secondary anti-rabbit (Jackson ImmunoResearch, 111-035-003, lot # 135002) diluted 1:10000 in 5% NFM in PBS for 1h at RT.
    • Wash membrane 2x for 5min with PBST and 2x for 5min with PBS.
    • Reveal protein bands using Perce ECL Western (Thermo Scientific, 32106, TE265714) with Amersham Hyperfilm ECL (GE Healthcare Limited, 28906835, 65901).
    Anmerkungen

    HEK293 cells with no endogenous PKP2 expression have been transfected with human FLAG-tagged PKP2 as a positive control. HL-1 cells express endogenous mouse PKP2.

  • 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
    Plakophilin 2 (PKP2)
    Andere Bezeichnung
    PKP2 (PKP2 Produkte)
    Synonyme
    ARVD9 antikoerper, 1200008D14Rik antikoerper, 1200012P04Rik antikoerper, AA516617 antikoerper, Pkp2l antikoerper, plakophilin 2 antikoerper, PKP2 antikoerper, Pkp2 antikoerper
    Hintergrund
    This gene encodes a member of the arm-repeat (armadillo) and plakophilin gene families. Plakophilin proteins contain numerous armadillo repeats, localize to cell desmosomes and nuclei, and participate in linking cadherins to intermediate filaments in the
    Alias Symbols: ARVD9
    Protein Interaction Partner: NDEL1, MTUS2, CEP250, DROSHA, BMI1, EED, RNF2, YWHAB, CLK2, YWHAE, WHSC1, UBC, SMAD9, YWHAQ, CUL3, MARK3, POLR3A, YWHAG, PKP2, JUP, KRT18, KRT5, DSG2, DSG1, DSP, DSC2, DSC1, CTNNB1, GTF2B, SFN,
    Protein Size: 837
    Molekulargewicht
    93 kDa
    Gen-ID
    5318
    NCBI Accession
    NM_001005242, NP_001005242
    UniProt
    A0AV37
    Pathways
    Cell-Cell Junction Organization, SARS-CoV-2 Protein Interaktom, Phosphorylierungen bei SARS-CoV-2 Infektion
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