电话:
400-7060-959
传真:
+86 10 56315212-8813
电子邮件:
orders@antibodies-online.cn

Phosphotyrosine 抗体

WB, IP, IF, IHC, ELISA, ICC 宿主: 兔 Polyclonal unconjugated
产品编号 ABIN361758
发货至: 中国
  • 抗原 See all Phosphotyrosine products
    Phosphotyrosine
    适用
    请咨询
    宿主
    • 90
    • 27
    • 1
    克隆类型
    • 102
    • 16
    多克隆
    标记
    • 41
    • 16
    • 10
    • 8
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This Phosphotyrosine antibody is un-conjugated
    应用范围
    • 84
    • 60
    • 39
    • 37
    • 36
    • 27
    • 23
    • 17
    • 13
    • 9
    • 9
    • 7
    • 6
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunoprecipitation (IP), Immunofluorescence (IF), Immunohistochemistry (IHC), ELISA, Immunocytochemistry (ICC)
    特异性
    Detects proteins phosphorylated on tyrosine residues. Does not cross-react with phosphoserine or threonine.
    纯化方法
    Peptide Affinity Purified
    免疫原
    Phosphotyrosine conjugated to KLH
  • 应用备注
    • WB (1:500)
    • IHC (1:100)
    • ICC/IF (1:50)
    • ELISA (1:2000)
    • IP (1:100)
    • optimal dilutions for assays should be determined by the user.
    说明

    A 1:250 dilution of ABIN361758 was sufficient for detection of tyrosine-phosphorylated species in mouse spleen lysates in western blot analysis.

    限制
    仅限研究用
  • 生效 #103692 (Proximity Ligation Assay)
    '独立验证'标志
    by
    Klinik für Anästhesie, Intensivmedizin und Schmerztherapie, Universitätsklinikum der Ruhr Universität Bochum
    No.
    #103692
    日期
    2019.04.29
    抗原
    Phosphotyrosine
    Lot Number
    1508290
    Method validated
    Proximity Ligation Assay
    Positive Control

    U937 monocytic cells stimulated with ODN1826

    Negative Control

    Anti pY antibody (ABIN361758) alone

    Anti TLR9 antibody (ABIN5542492) alone

    Notes

    Passed. PLA signal visible in stimulated cells. Negative control with only one primary antibody shows no signal.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN361758
    Secondary Antibody
    ABIN5542492
    Full Protocol
    • Grow U937 cells in RPMI (Gibco, 21875-034) supplemented with 10% FCS (Gibco, A3840001) and Pen/Strep(Gibco, 15140-122), at 37°C and 5% CO2 in 500µL in 12-well plates.
    • Incubate the cells with 1µM ODN1826 for 4h.
    • Use cytospin to spin cells onto slide.
    • Fix cells on coverslide in 4% PFA for 20min at RT.
    • Wash cells 3x for 5min with PBS.
    • Permeabilize cells in PBS containing 0.1% Triton for 5min at RT.
    • Block non-specific binding with Duolink PLA Blocking Solution (Sigma-Aldrich) for 30min at RT.
    • Incubate cells with primary rabbit anti-pY antibody (antibodies-online, ABIN361758, lot 1508290) diluted 1:100 and primary mouse anti-TLR9 antibody (antibodies-online, ABIN5542492, lot 160127) diluted 1:200 ON at 4°C.
    • Wash cells 3x for 5min with PBS.
    • Incubate cells with Duolink In Situ PLA Probe anti-Rabbit MINUS (Sigma-Aldrich, DUO92005, lot SLBZ4516) and Duolink In Situ PLA Probe anti-Mouse mouse PLUS (Sigma-Aldrich, DUO92001, lot SLBZ8369) proximity probes according to manufacturers recommendations.
    • Wash cells 2x for 5min with PBS.
    • Perform ligation step according to the Duolink PLA kit (Sigma).
    • Perform amplification step according to the Duolink In Situ PlA Detection Kit Green (Sigma-Aldrich, DUO92014).
    • Mount coverslips on glass slides in ProLong Gold antifade reagent (Invitrogen, P36935, lot 1890418) containing DAPI.
    • Image acquisition with an Olympus widefield microscope.
    Experimental Notes

    We used double the amount the Phi29 polymerase for the amplifaction step than recommended.

  • 状态
    Liquid
    浓度
    0.25 mg/mL
    缓冲液
    PBS pH 7.0, 50 % glycerol, 0.01 % sodium azide, Storage buffer may change when conjugated
    储存液
    Sodium azide
    注意事项
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    储存条件
    -20 °C
    储存方法
    -20°C
  • 抗原
    Phosphotyrosine
    Abstract
    Phosphotyrosine 产品
    物质类
    Amino Acid
    背景
    Protein phosphorylation is an important posttranslational modification that serves many key functions to regulate a protein's activity, localization, and protein-protein interactions. Phosphorylation is catalyzed by various specific protein kinases, which involves removing a phosphate group from ATP and covalently attaching it to to a recipient protein that acts as a substrate. Most kinases act on both serine and threonine, others act on tyrosine, and a number (dual specificity kinases) act on all three. Because phosphorylation can occur at multiple sites on any given protein, it can therefore change the function or localization of that protein at any time (1). Changing the function of these proteins has been linked to a number of diseases, including cancer, diabetes, heart disease, inflammation and neurological disorders (2-4). In particular, the phosphorylation of tyrosine is considered one of the key steps in signal transduction and regulation of enzymatic activity (5). Phosphotyrosine can be detected through specific antibodies, and are helpful in facilitating the identification of tyrosine kinase substrates (6).
You are here:
客服