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c-MYC 抗体 (AA 31-80)

This anti-c-MYC antibody is a 兔 多克隆 antibody detecting c-MYC in WB, IHC, ELISA 和 IF. Suitable for 人, 小鼠 和 大鼠. Independently validated for use in Western Blotting, Immunocytochemistry.
产品编号 ABIN1532205
发货至: 中国

Quick Overview for c-MYC 抗体 (AA 31-80) (ABIN1532205)

抗原

See all c-MYC (MYC) 抗体
c-MYC (MYC) (Myc Proto-Oncogene protein (MYC))

适用

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人, 小鼠, 大鼠

宿主

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克隆类型

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多克隆

标记

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This c-MYC antibody is un-conjugated

应用范围

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Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Immunofluorescence (IF)
  • 抗原表位

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    AA 31-80

    特异性

    MYC Antibody detects endogenous levels of total MYC protein.

    纯化方法

    The antibody was purified from rabbit antiserum by affinity-chromatography using immunogen.

    纯度

    > 95 %

    免疫原

    The antiserum was produced against synthesized peptide derived from human MYC.

    亚型

    IgG
  • 应用备注

    WB: 1:500~1:1000 IHC: 1:50~1:100 IF: 1:100~1:500 ELISA: 1:40000

    说明

    Unigene-Number: Hs.202453 (NCBI Gene Symbol: MYC)

    限制

    仅限研究用
  • 生效 #102753 (Western Blotting)
    '独立验证'标志
    by
    Developmental Biology, Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Georg-August-University of Göttingen
    No.
    #102753
    日期
    2018.07.03
    抗原
    MYC
    Lot Number
    210020 and 424170020
    Method validated
    Western Blotting
    Positive Control

    HEK-293 cells transfected with Myc expression plasmid

    Negative Control

    HEK-293 cells transfected with HA-Spag4-myc expression plasmid

    Notes

    Passed. ABIN1532205 detects Myc protein in a western blot. No cross-reaction was observed with the myc-tag sequence positioned at the C-terminus of a recombinant protein.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1532205
    Secondary Antibody
    rabbit anti-mouse IgG, HRP-linked (Sigma A9044, Lot 034M4761)
    Full Protocol
    • Grow HEK-293 cells (ATCC, CRL-1658) in DMEM+GlutaMAXDMEM+GlutaMAX (Gibco, 10567-014, lot 1922818) supplemented with fetal bovine serum (Gibco, 10500-064, lot 08FO477K) and Pen/Strep (Gibco, 15140-122, lot 1924797) at 37°C and 5% CO2 to 70% confluency.
    • Transfect cells either with a Myc expression plasmid (pcDNA3.1-Myc) or HA- and myc-tagged SPAG4 expression plasmids using EndofectinMax (GeneCopoeia) following the manufacturer´s instructions.
    • Grow cells for 24h.
    • Lyse cells in SDS-sample buffer and denature total cellular lysates for 5min at 95°C. Subsequently separate them on a denaturing 10% SDS-PAGE (Laemmli 1970).
    • Transfer proteins onto 0.2µm Protran membrane (GE Healthcare, 10600004, A10043108) by wet blotting for 1h at 400mA (Towbin et al., 1979).
    • Block the membrane in TBST (50mM Tris-HCl, pH7.4, 150mM NaCl, 0.2% Tween 20) containing 5% milk (blocking solution) for 60min at RT.
    • Incubate membrane with primary rabbit anti-myc antibody (antibodies-online, ABIN1532205 Lot#424170020 and Lot#210020) diluted 1:500 in blocking solution overnight at 4°C.
    • Wash membrane with TBST for 45min at RT.
    • Incubate membrane with secondary goat anti-rabbit IgG (H+L), HRP-linked (Jackson Immuno Research, 111-035-003, lot 123450) diluted 1:1000 in blocking solution for 45min at RT.
    • Wash membrane 6x for 5min with TBST.
    • Reveal protein bands using Clarity Max Western ECL substrate (Bio-Rad, 1705062) and capture images via Chemidoc Imaging System (BioRad).
    • Incubate membrane with primary mouse anti-HA-tag antibody (clone 12CA5) diluted 1:500 in blocking solution overnight at 4°C.
    • Wash membrane with TBST for 45min at RT.
    • Incubate membrane with secondary rabbit anti-mouse IgG, HRP-linked (Sigma A9044, Lot 034M4761) diluted 1:1000 in blocking solution for 45min at RT.
    • Reveal protein bands using Clarity Max Western ECL substrate (Bio-Rad, 1705062).
    Experimental Notes

    In lysates of HEK-293 cells ectopically expressing recombinant protein with an N-terminal HA- and a C-terminal myc-tag ABIN1532205 did not detect the labeled protein at the expected molecular weight. In contrast, an anti-HA-tag antibody (clone 12CA5) revealed a protein at the expected MW.

  • 生效 #103023 (Immunocytochemistry)
    '独立验证'标志
    by
    Developmental Biology, Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Georg-August-University of Göttingen
    No.
    #103023
    日期
    2018.07.03
    抗原
    MYC
    Lot Number
    210020 and 424170020
    Method validated
    Immunocytochemistry
    Positive Control

    HEK-293 cells transfected with Myc expression plasmid

    Negative Control

    HEK-293 cells transfected with HA-Spag4-myc expression plasmid

    Notes

    Passed. ABIN1532205 (lot 424170020 and lot 210020) detects Myc in immunocytochemistry. No cross-reaction was observed with the myc-tag sequence positioned at the C-terminus of a recombinant protein.

    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1532205
    Secondary Antibody
    goat anti-rabbit IgG MFP590 (Mobitec MFP-A1037, lot 3002088), F(ab')2-Goat anti-Rabbit IgG (H+L) Alexa Fluor 555 (Life Technologies, A21430, lot 1637266)
    Full Protocol
    • Grow HEK-293 cells (ATCC, CRL-1658) in DMEM+GlutaMAXDMEM+GlutaMAX (Gibco, 10567-014, lot 1922818) supplemented with fetal bovine serum (Gibco, 10500-064, lot 08FO477K) and Pen/Strep (Gibco, 15140-122, lot 1924797) at 37°C and 5% CO2 to 70% confluency.
    • Transfect cells either with a Myc expression plasmid (pcDNA3.1-Myc) or HA- and myc-tagged SPAG4 expression plasmids using EndofectinMax (GeneCopoeia) following the manufacturer´s instructions.
    • Grow cells for 24h.
    • Fix cells in 3.7% paraformaldehyde (in PBS) for 20min at 4°C followed by incubation in 0.3% Triton X-100 for 10min at 4°C.
    • Block cells in PBS containing 1% BSA and 0.5% Tween-20 (PBT) for 1h at RT.
    • Incubate cells with primary antibody
      • rabbit anti-Myc antibody (antibodies-online, ABIN1532205, lot 424170020 or lot 210020) diluted 1:50 in PBS and mouse anti-HA tag antibody (clone 12CA5) ON at 4°C (images G, H).
      • rabbit anti-Myc antibody (antibodies-online, ABIN1532205, lot 210020 or lot 210020) diluted 1:50 in PBS ON at 4°C (images A, D).
    • Wash cells with 50mM Tris-HCl, pH7.4, 150mM NaCl, 0.1% Tween 20 (TBST) for 15min at RT.
    • Incubate cells with secondary antibody
      • goat anti-rabbit IgG MFP590 (Mobitec MFP-A1037, lot 3002088) diluted 1:300 in PBS and goat anti-mouse IgGDylight488 (Thermo Scientific, 35503) diluted 1:10000 in PBS for 1h at 37°C (images G, H).
      • F(ab')2-Goat anti-Rabbit IgG (H+L) Alexa Fluor 555 (Life Technologies, A21430, lot 1637266) diluted 1:2000 in PBS for 45min at 37°C (images A, D).
    • Counterstain cells with DAPI (Sigma, D-9542).
    • Image acquisition on Zeiss LSM 510 confocal microscope and processing using Adobe Photoshop 5.0.
    Experimental Notes

    The antibody detects Myc proteins by immunoblotting and by immunocytology. No cross-reaction with the C-terminal Myc-tag.

  • 状态

    Liquid

    浓度

    1 mg/mL

    缓冲液

    phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150 mM NaCl, 0.02 % sodium azide and 50 % glycerol.

    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

    Stable at -20°C for at least 1 year.

    有效期

    12 months
  • 抗原

    c-MYC (MYC) (Myc Proto-Oncogene protein (MYC))

    别名

    Myc Proto-Oncogene protein

    背景

    Synonyms: Myc proto-oncogene protein, Class E basic helix-loop-helix protein 39, bHLHe39, Proto-oncogene c-Myc, Transcription factor p64 , MYC , BHLHE39

    NCBI Gene Symbol: MYC

    分子量

    48 kDa

    基因ID

    4609

    OMIM

    113970

    UniProt

    P01106

    途径

    p53 Pathway, Cell Division Cycle, Sensory Perception of Sound, Transition Metal Ion Homeostasis, Mitotic G1-G1/S Phases, Positive Regulation of Endopeptidase Activity, Regulation of Carbohydrate Metabolic Process, Positive Regulation of Response to DNA Damage Stimulus, Warburg Effect
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