Plakophilin 2 抗体 (AA 801-881)
(1 reference)
(1 validation)-
- 抗原 See all Plakophilin 2 (PKP2) 抗体
- Plakophilin 2 (PKP2)
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抗原表位
- AA 801-881
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适用
- 人
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- This Plakophilin 2 antibody is un-conjugated
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应用范围
- ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p))
- 交叉反应
- 人
- 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
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anti-Plakophilin 2 (PKP2) (N-Term) antibody
PKP2 适用: 人, 大鼠, 小鼠, Cow, 犬, 马 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) (AA 72-194) antibodyPKP2 适用: 人 WB, IHC (p), IF 宿主: 兔 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) (C-Term) antibodyPKP2 适用: 人 WB, ELISA 宿主: 兔 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) (AA 61-183) antibodyPKP2 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) (C-Term) antibodyPKP2 适用: 人, 大鼠 WB, ELISA 宿主: 兔 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) (AA 1-350) antibodyPKP2 适用: 人, 小鼠 WB, IHC (p), IP, FACS 宿主: 小鼠 Monoclonal 8H6 unconjugated
anti-Plakophilin 2 (PKP2) (N-Term) antibodyPKP2 适用: 人, 大鼠 WB, ELISA, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) (AA 125-139) antibodyVerified PKP2 适用: 人, 小鼠 WB, ELISA, IHC 宿主: 山羊 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) (AA 571-849) antibodyPKP2 适用: 人 WB, IHC, ICC 宿主: 兔 Polyclonal unconjugated
anti-Plakophilin 2 (PKP2) antibodyPKP2 适用: 人, 大鼠, 小鼠 WB, IHC, IF 宿主: 兔 Polyclonal unconjugated
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- 应用备注
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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
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- 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.
- 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).
生效 #103656 (Immunofluorescence)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![A. IF staining of cells expressing PKP2-FLAG using ABIN1386389 (green) or an anti-FLAG-tag antibody (red). Nuclear staining with DAPI (blue) (magnification 60x). B. Staining of HeLa cells not expressing PKP2-FLAG using ABIN1386389 (magnification 40x). C. Staining of cells expressing PKP2-Flag with a secondary antibody only (magnification 40x). D and E. Staining of endogenous Pkp2 in mouse heart tissue with ABIN1386389 (D) or secondary antibody only (E) (magnification 40x).]()
A. IF staining of cells expressing PKP2-FLAG using ABIN1386389 (green) or an anti-FLAG-tag antibody (red). Nuclear staining with DAPI (blue) (magnification 60x). B. Staining of HeLa cells not expressing PKP2-FLAG using ABIN1386389 (magnification 40x). C. Staining of cells expressing PKP2-Flag with a secondary antibody only (magnification 40x). D and E. Staining of endogenous Pkp2 in mouse heart tissue with ABIN1386389 (D) or secondary antibody only (E) (magnification 40x).
Full Methods -
- 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
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: "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).
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: "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).
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- 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
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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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