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Plakophilin 2 抗体 (AA 801-881)

PKP2 适用: 人 ELISA, IHC (p), ICC, IHC (fro), IF (p), IF (cc) 宿主: 兔 Polyclonal unconjugated
产品编号 ABIN1386389
发货至: 中国
  • 抗原 See all Plakophilin 2 (PKP2) 抗体
    Plakophilin 2 (PKP2)
    抗原表位
    • 14
    • 8
    • 7
    • 5
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    AA 801-881
    适用
    • 47
    • 18
    • 18
    • 3
    • 3
    • 3
    • 1
    • 1
    • 1
    • 1
    • 1
    宿主
    • 42
    • 3
    • 1
    • 1
    克隆类型
    • 44
    • 3
    多克隆
    标记
    • 25
    • 4
    • 4
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This Plakophilin 2 antibody is un-conjugated
    应用范围
    • 35
    • 22
    • 21
    • 4
    • 3
    • 3
    • 2
    • 1
    • 1
    ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunofluorescence (Cultured Cells) (IF (cc))
    交叉反应
    预测反应
    Mouse,Rat,Dog,Cow,Sheep,Pig,Horse,Rabbit
    纯化方法
    Purified by Protein A.
    免疫原
    KLH conjugated synthetic peptide derived from human Plakophilin 2
    亚型
    IgG
  • 应用备注
    ELISA 1:500-1000
    IHC-P 1:200-400
    IHC-F 1:100-500
    IF(IHC-P) 1:50-200
    IF(IHC-F) 1:50-200
    IF(ICC) 1:50-200
    ICC 1:100-500
    限制
    仅限研究用
  • 生效 #103656 (Immunofluorescence)
    '独立验证'标志
    by
    Gencardio (IDIBGI)
    No.
    #103656
    日期
    2019.03.13
    抗原
    PKP2
    Lot Number
    AD102900
    Method validated
    Immunofluorescence
    Positive Control

    HL1 cells

    HeLa cells

    mouse heart tissue

    Negative Control

    HL1 cells

    HeLa cells

    secondary antibody only control

    Notes

    ABIN1386389 specifically labels the targeted antigen in mouse heart. No signal was detected in sample negative control and the secondary antibody only control.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1386389
    Secondary Antibody
    goat anti-rabbit AF488 antibody (Invitrogen, A11008, 1911238)
    Full Protocol
    • HL1 and HeLa cells
      • Grow HL1 cells in Claycomb medium (sigma, 51800C, SLBX8700) supplemented with 10% FBS (Gibco, 10270-106, 42G5178K), 1% Penicillin-Streptomycin (Sigma, P4333, lot 048M4874V) and 1% Glutamax (Gibco, 35050-061, lot 2037045), at 37°C and 5% CO2 in 12mm coverslips in a well of 12 well plate.
      • Grow HeLa cells in MEM/EBSS medium (GE Life Science, SH30244.01, lot AC14563283) supplemented with 10% FBS (Gibco, 10270-106, lot 42G5178K), 1% Penicillin-Streptomycin (Sigma, P4333, lot 048M4874V) and 1% Glutamax (Gibco, 35050-061, lot 2037045), at 37°C and 5% CO2 in 12mm coverslips in a well of 12 well plate.
      • Transfect HeLa cells with PKP2a plasmid (p915 PKP2a-pFLAG-cmv5a, provided by Dr. Green KJ) using Lipofectamine 3000 (Invitrogen, L3000-015, lot 2041107) following the manufacturer´s instructions.
      • Wash cells 1x with PBS.
      • Fix cells on coverslips in 4% PFA for 20min at RT.
      • Wash cells 3x for 5min with PBS.
      • Incubate coverslips with quench solution (PBS with 75mM glycine and 20mM NH4Cl) for 10min at RT.
      • Wash cells 1x with PBS.
      • Permeabilize cells and block non-specific binding with PFS-like buffer containing 0.1% BSA, 0.1% triton and 0.02% NaN3 for 30min at 37°C.
      • Incubate cells with primary anti-Plakophilin 2 (PKP2) (AA 831-881) antibody (antibodies-online, ABIN1386389, lot AD102900) diluted 1:60 in PFS-like buffer or with anti-Flag-tag antibody (Sigma, monoclonal anti-flag, F316, lot SLBT7654) diluted 1:400 in PFS-like buffer ON at 4°C.
      • Wash cells 3x for 5min with PBS.
      • Incubate cells with secondary goat anti-rabbit AF488 antibody (Invitrogen, A11008, lot 1911238) or with secondary goat anti-mouse AF568 antibody (Invitrogen, A11031, lot 1841757) diluted 1:200 in PFS-like buffer for 30min at RT.
      • Wash cells 2x for 5min with PFS-like buffer.
      • Wash cells 2x for 5min with PBS.
      • DAPI counterstain for 5min at RT.
      • Mount coverslips on glass slides in Calbicochem (Millipore, 345789, lot 2682585).
      • Image acquisition with NIS-ELEMENTS (Nikon).
    • Tissue: mouse heart
      • Obtain mouse heart mouse and wash it with PBS.
      • Fix tissue in 15ml falcon tube with 4% PFA ON at 4°C.
      • Wash the fixed heart with water.
      • Embed tissue in paraffin.
      • Cut the tissue using a microtrom and put it into slides.
      • Desparaffinize the tissue in the slide (from xylene to water).
      • Incubate the slides with retrieval solution for 30min at 95°C to permeabilize the tissue.
      • Cool slides down on ice.
      • Wash tissue 2x for 3min with PBS.
      • Wash tissue 2x for 3min with PBS 0.1% triton.
      • Block non-specific binding with PBS 0.1% triton 10% BSA for 1h at RT.
      • Incubate tissue with primary anti-PKP2 (AA 831-881) antibody (antibodies-online, ABIN1386389, lot AD102900) diluted 1:100 in PBS 1% BSA solution ON at 4°C.
      • Wash tissue 2x for 5min with PBS 0.1% triton.
      • Incubate tissue with secondary goat anti-rabbit AF488 antibody (Invitrogen, A11008, 1911238) diluted 1:200 in PBS 1% BSA solution for 1h at RT.
      • Wash cells 2x for 5min with PBS.
      • Wash tissue 2x for 3min with PBS 0.1% triton.
      • Wash tissue 2x for 3min with PBS.
      • DAPI counterstain for 5min at RT.
      • Mount slides with coverslips in Calbicochem (Millipore, 345789, lot 2682585).
      • Image acquisition with NIS-ELEMENTS (Nikon).
    Experimental Notes

    ABIN1386389 did not label recombinantly expressed PKP2-FLAG in transfected HL1 cells. A hypothesis could be that antibody recognizes the murine protein in mouse heart samples but not the human sequence of the vector (transfected HL1).

  • 状态
    Liquid
    浓度
    1 μg/μL
    缓冲液
    0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.
    储存液
    ProClin
    注意事项
    This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
    储存条件
    4 °C,-20 °C
    储存方法
    Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
    有效期
    12 months
  • Akdis, Medeiros-Domingo, Gaertner-Rommel, Kast, Enseleit, Bode, Klingel, Kandolf, Renois, Andreoletti, Akdis, Milting, Lüscher, Brunckhorst, Saguner, Duru: "Myocardial expression profiles of candidate molecules in patients with arrhythmogenic right ventricular cardiomyopathy/dysplasia compared to those with dilated cardiomyopathy and healthy controls." in: Heart rhythm, Vol. 13, Issue 3, pp. 731-41, (2016) (PubMed).

  • 抗原
    Plakophilin 2 (PKP2)
    别名
    Plakophilin 2 (PKP2 产品)
    别名
    ARVD9 antibody, 1200008D14Rik antibody, 1200012P04Rik antibody, AA516617 antibody, Pkp2l antibody, plakophilin 2 antibody, PKP2 antibody, Pkp2 antibody
    背景

    Synonyms: ARVD 9, ARVD-9, ARVD9, PKP 2, PKP2, PKP-2, Plakophilin-2. Plakophilin2, PKP2_HUMAN.

    Background: Plakophilins 1, 2, 3 and 4 (PKP1-4) influence development and participate in linking cadherins to cytoskeletal intermediate filaments. Plakophilins 1-4 contain arm-repeat (armadillo) domains, and localize to nuclei and cell desmosomes (cell-cell junctions found in suprabasal layers of stratifying epithelia that undergo mechanical stress). Plakophilin-1 mediates increases in desmosomal protein content, desmosome assembly, and regulation of cell migration. Plakophilin-2 is important for desmosome assembly and is an essential morphogenic factor and architectural component of the heart. Plakophilin-3 plays a role in both desmosome-dependent adhesion and signaling pathways. Plakophilin-4 is a component of desmosomal adhesion plaques that regulates junctional plaque organization and cadherin function.

    基因ID
    5318
    UniProt
    Q99959
    途径
    Cell-Cell Junction Organization, SARS-CoV-2 Protein Interactome, The Global Phosphorylation Landscape of SARS-CoV-2 Infection
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