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Epigenetics and Nuclear Signaling Transcription Domain Families HLH / Leucine Zipper HLH / Leucine Zipper
使用敲除细胞株进行验证重组RabMAb

重组Anti-c-Myc抗体[Y69] (ab32072)

  • Datasheet
  • SDS
Reviews (50)Q&A (8)References (602)

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Western blot - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunoprecipitation - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunocytochemistry - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
  • Flow Cytometry - Anti-c-Myc antibody [Y69] (ab32072)
  • OI-RD Scanning - Anti-c-Myc antibody [Y69] (ab32072)
  • Anti-c-Myc antibody [Y69] (ab32072)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y69] to c-Myc
  • Suitable for: WB, ICC/IF, Flow Cyt, IHC-P, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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概述

  • 产品名称

    Anti-c-Myc抗体[Y69]
    参阅全部 c-Myc 一抗
  • 描述

    兔单克隆抗体[Y69] to c-Myc
  • 宿主

    Rabbit
  • 特异性

    This antibody is specific for endogenous c-Myc. It does not detect Myc tag. Expression levels of the target protein vary with sample type and some optimization may be required.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • 免疫原

    Synthetic peptide within Human c-Myc aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P01106
    (Peptide available as ab166837)

  • 阳性对照

    • 购买相配的WB阳性对照Recombinant human c-Myc protein (Active)
    • WB: Jurkat, HeLa, HEK-293T, Raji, MCF-7, K562, THP1, A20, AR42J, Rat-1, rat spleen and pancreas, L6, Neuro-2a, Raw264.7 cell lysates, L363 MM and CA46 cells. ICC/IF: HEK293 and HeLa cells. IHC-P: Human Burkitt lymphoma, diffuse large B cell lymphoma, adenocarcinoma of the colon, lung adenocarcinoma, gastric adenocarcinoma, urinary bladder transitional carcinoma, esophagus, glioblastoma and low-grade glioma tumor tissues. IP: Jurkat whole cell lysate (ab7899). Flow Cyt: HeLa cells.
  • 常规说明

    The proto-oncogene MYC plays a role in human oncogenesis. For more information see here. 

    This recombinant rabbit monoclonal antibody (Y69) to c-Myc specifically detects endogenous c-Myc. ab32, the Mouse monoclonal antibody (9E10 clone) to the Myc tag however can be used to study Myc-tagged proteins. More information comparing the two antibodies can be found here.

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 解离常数(KD)

    KD = 3.80 x 10 -12 M
    Learn more about KD
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    Y69
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • HLH / Leucine Zipper
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Nuclear
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Cell cycle
    • Cell differentiation
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Transcription factors
    • Cancer
    • Tumor biomarkers
    • Oncoproteins
    • Signal Transduction
    • Antibodies
    • c-myc

相关产品

  • Alternative Versions

    • Anti-c-Myc antibody [Y69] - BSA and Azide free (ab168727)
    • Alexa Fluor® 488 Anti-c-Myc antibody [Y69] (ab190026)
    • Alexa Fluor® 647 Anti-c-Myc antibody [Y69] (ab190560)
    • Alexa Fluor® 594 Anti-c-Myc antibody [Y69] (ab201775)
    • Alexa Fluor® 555 Anti-c-Myc antibody [Y69] (ab201780)
    • HRP Anti-c-Myc antibody [Y69] (ab205818)
    • FITC Anti-c-Myc antibody [Y69] (ab223913)
  • Assay kits

    • c-Myc Transcription Factor Assay Kit (Colorimetric) (ab207200)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Immunizing Peptide (Blocking)

    • Human c-Myc peptide (ab166837)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human MYC (c-Myc) knockout HEK-293T cell line (ab256500)
  • KO cell lysates

    • Human MYC (c-Myc) knockout HEK-293T cell lysate (ab263850)
  • KO cell pellets

    • Human MYC (c-Myc) knockout HEK293T cell pellet (ab278810)
  • Positive Controls

    • Raji whole cell lysate (ab30124)
    • Jurkat whole cell lysate (ab30128)
    • A20 whole cell lysate (ab7180)
    • Jurkat whole cell lysate (ab7899)
    • K-562 whole cell lysate (ab7911)
    • THP-1 whole cell lysate (ab7913)
  • Recombinant Protein

    • Recombinant human c-Myc protein (Active) (ab169901)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • ECL Substrate Kit (High Sensitivity) (ab133406)
    • Anti-PCNA antibody [PC10] (ab29)
    • Anti-Myc tag antibody [9E10] (ab32)
    • Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker (ab6326)
    • Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)

应用

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32072 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
Flow Cyt
Human
ICC
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Mouse
Rat
Human
应用 Ab评论 说明
WB (17)
1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 49 kDa).Can be blocked with Human c-Myc peptide (ab166837).
ICC/IF (7)
Use a concentration of 5 - 10 µg/ml.

1/100.

Flow Cyt
1/76.
IHC-P (16)
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP (6)
Use a concentration of 5 µg/ml.
说明
WB
1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 49 kDa).Can be blocked with Human c-Myc peptide (ab166837).
ICC/IF
Use a concentration of 5 - 10 µg/ml.

1/100.

Flow Cyt
1/76.
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP
Use a concentration of 5 µg/ml.

靶标

  • 功能

    Participates in the regulation of gene transcription. Binds DNA in a non-specific manner, yet also specifically recognizes the core sequence 5'-CAC[GA]TG-3'. Seems to activate the transcription of growth-related genes.
  • 疾病相关

    Note=Overexpression of MYC is implicated in the etiology of a variety of hematopoietic tumors.
    Note=A chromosomal aberration involving MYC may be a cause of a form of B-cell chronic lymphocytic leukemia. Translocation t(8;12)(q24;q22) with BTG1.
    Defects in MYC are a cause of Burkitt lymphoma (BL) [MIM:113970]. A form of undifferentiated malignant lymphoma commonly manifested as a large osteolytic lesion in the jaw or as an abdominal mass. Note=Chromosomal aberrations involving MYC are usually found in Burkitt lymphoma. Translocations t(8;14), t(8;22) or t(2;8) which juxtapose MYC to one of the heavy or light chain immunoglobulin gene loci.
  • 序列相似性

    Contains 1 basic helix-loop-helix (bHLH) domain.
  • 翻译后修饰

    Phosphorylated by PRKDC. Phosphorylation at Thr-58 and Ser-62 by GSK3 is required for ubiquitination and degradation by the proteasome.
    Ubiquitinated by the SCF(FBXW7) complex when phosphorylated at Thr-58 and Ser-62, leading to its degradation by the proteasome. In the nucleoplasm, ubiquitination is counteracted by USP28, which interacts with isoform 1 of FBXW7 (FBW7alpha), leading to its deubiquitination and preventing degradation. In the nucleolus, however, ubiquitination is not counteracted by USP28, due to the lack of interaction between isoform 4 of FBXW7 (FBW7gamma) and USP28, explaining the selective MYC degradation in the nucleolus. Also polyubiquitinated by the DCX(TRUSS) complex.
  • 细胞定位

    Nucleus > nucleoplasm. Nucleus > nucleolus.
  • Target information above from: UniProt accession P01106 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 4609 Human
    • Entrez Gene: 17869 Mouse
    • Entrez Gene: 24577 Rat
    • Omim: 190080 Human
    • SwissProt: P01106 Human
    • SwissProt: P01108 Mouse
    • SwissProt: P09416 Rat
    • Unigene: 202453 Human
    • Unigene: 2444 Mouse
    • Unigene: 12072 Rat
    see all
  • 形式

    c-Myc is also expressed in the cytoplasm.
  • 别名

    • AU016757 antibody
    • Avian myelocytomatosis viral oncogene homolog antibody
    • bHLHe39 antibody
    • c Myc antibody
    • Cellular myelocytomatosis oncogene antibody
    • Class E basic helix-loop-helix protein 39 antibody
    • MGC105490 antibody
    • MRTL antibody
    • Myc antibody
    • Myc protein antibody
    • Myc proto oncogene protein antibody
    • Myc proto-oncogene protein antibody
    • myc-related translation/localization regulatory factor antibody
    • MYC_HUMAN antibody
    • Myc2 antibody
    • myca antibody
    • MYCC antibody
    • Myelocytomatosis oncogene a antibody
    • Myelocytomatosis oncogene antibody
    • Niard antibody
    • Nird antibody
    • oncogene c-Myc antibody
    • Oncogene Myc antibody
    • OTTHUMP00000158589 antibody
    • OTTHUMP00000227763 antibody
    • Proto-oncogene c-Myc antibody
    • Protooncogene homologous to myelocytomatosis virus antibody
    • RNCMYC antibody
    • Transcription factor p64 antibody
    • Transcriptional regulator Myc-A antibody
    • V-Myc avian myelocytomatosis viral oncogene homolog antibody
    • v-myc myelocytomatosis viral oncogene homolog (avian) antibody
    • zc-myc antibody
    see all

图片

  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/1000 dilution

    Lane 1 : Wild-type Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Wild-type HEK-293T cell lysate
    Lane 4 : MYC knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 57 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab32072 observed at 57 kDa. Red - loading control, ab8245 observed at 37 kDa.

     ab32072 was shown to react with MYC in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab256500 (knockout cell lysate ab263850) was used. Wild-type and MYC knockout samples were subjected to SDS-PAGE. ab32072 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Immunoprecipitation - Anti-c-Myc antibody [Y69] (ab32072)
    Immunoprecipitation - Anti-c-Myc antibody [Y69] (ab32072)

    c-Myc was immunoprecipitated using 0.5mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell extract, 5µg of unpurified rabbit monoclonal to c-Myc [Y69] and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with unpurified ab32072.

    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 57kDa; c-Myc [Y69]

  • Immunocytochemistry - Anti-c-Myc antibody [Y69] (ab32072)
    Immunocytochemistry - Anti-c-Myc antibody [Y69] (ab32072)
    ab32072 staining MYC in wild-type HEK293 cells (top panel) and MYC knockout HEK293 cells (ab256500) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32072 at 5μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)Image from Kluk MJ et al., PLoS One. 2012;7(4):e33813. Fig 1.; doi: 10.1371/journal.pone.0033813. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Photomicrographs of select tumors and reactive tissue stained for c-Myc (positive staining  is the brown nuclei). Positive control (Burkitt lymphoma with a confirmed c-Myc translocation) revealed uniform, intense staining in >90% of tumor cells (Burkitt). In contrast, reactive lymphoid tissue revealed variable staining in only 10% of normal lymphocyte nuclei (Tonsil). Representative images from Diffuse large B cell lymphoma (DLBCL) cases and associated percent c-Myc+ tumor nuclei: Case 1, 90% MYC+; Case 7, 70% MYC+; and Cases 35 and 38, 30% c-Myc+. c-Myc staining was exclusively nuclear in all cases under the described staining conditions.

  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/1000 dilution

    Lane 1 : Rat pancreas lysates
    Lane 2 : AR42J (Rat pancreatic tumor epithelial cell) whole cell lysates
    Lane 3 : Rat-1 (Rat embryonic fibroblast) whole cell lysates
    Lane 4 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 5 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 20000 µg/ml

    Predicted band size: 49 kDa



    Blocking/Diluting buffer and concentation: 5% NFDM/TBST

    Lanes 1 and 2:  80 seconds exposure time 

    Lanes 3 to 5: 5 seconds exposure time 

    Observed MW: 57 KDa

  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    Western blot - Anti-c-Myc antibody [Y69] (ab32072)This image is taken from Calabrese D. R. et al. Nat Commun. 2018; 9: 4229. Fig 3e doi: 10.1038/s41467-018-06315-w Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

    Western blot of L363 MM cell and CA46 cells measuring expresison of c-Myc (using ab32072) and GAPDH as the dose of DC-34 increases. 

    c-Myc protein levels are inhibited as a function of the dose of DC-34 in L363 cells; only the highest dose of DC-34 affected c-Myc in the more resistant CA46 Burkitt’s lymphoma cells.

  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
    Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)

    ab32072 staining c-Myc in HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32072 at 10μg/ml dilution (shown in green) and ab195889, mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)Image from Toon CW et al., PLoS One. 2014;9(2):e87456. Fig 1.; doi: 10.1371/journal.pone.0087456. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Human colorectal carcinoma (CRC) tissues stained for c-Myc using ab32072 at 1/100 dilution in immunohistochemical analysis.

    Panel A: c-Myc positive IHC staining.

    Panel B: c-Myc negative IHC staining.

    For the full image see PMID 24503701.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)

    IHC image of ab32072 staining c-Myc in human adenocarcinoma formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32072, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)

    IHC image of ab32072 staining c-Myc in human esophagus formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32072, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the Secondary only control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)Image from Simeone P et al., PLoS One. 2014;;9(7):e103030.; doi: 10.1371/journal.pone.0103030. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Expression of c-Myc, as determined by immunohistochemical staining of glioblastoma sample (left) and low-grade glioma tumor (right) with ab32072. Representative samples are shown. Scale bars  = 20 µm. Nuclei were counterstained with hematoxylin (in blue).

    For the full image see PMID 25050814.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human diffuse large B cell lymphoma tissue labelling c-Myc with purified ab32072 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling c-Myc with purified ab32072 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarinoma of colon tissue labelling c-Myc with unpurified ab32072.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-c-Myc antibody [Y69] (ab32072)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarinoma tissue labelling c-Myc with unpurified ab32072.

  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/1000 dilution (unpurified)

    Lane 1 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 2 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
    Lane 3 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate
    Lane 4 : A20 (Mouse B lymphoma cell line) Whole Cell Lysate
    Lane 5 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 57 kDa why is the actual band size different from the predicted?



    The predicted molecular weight of c-Myc is 48 kDa (SwissProt), however we expect to observe a banding pattern at 57 kDa.

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab32072 overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP antibody, and visualised using ECL development solution ab133406.

  • Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    Western blot - Anti-c-Myc antibody [Y69] (ab32072)
    All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/1000 dilution

    Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates
    Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
    Lane 4 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
    Lane 5 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates
    Lane 6 : Rat spleen whole cell lysates
    Lane 7 : L6 (Rat skeletal muscle myoblast) whole cell lysates
    Lane 8 : Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates
    Lane 9 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size: 49 kDa
    Observed band size: 57 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
    Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)

    Immunocytochemistry/immunofluorescence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling c-Myc with purified ab32072 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) (1/500).

  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
    Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)Image courtesy of Dr Vladimir Milenkovic by Abreview.

    Unpurified ab32072 staining c-Myc in HEK293 cells transfected with CACNB4-c-Myc by immunocytochemistry/ immunofluorescence.Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100 then blocked using 5% serum for 20 minutes at 25°C. Samples were then incubated with ab32072 at a 1/250 dilution for 16 hours at 4°C. The secondary used was an Alexa Fluor® 488 conjugated goat anti-rabbit polyclonal, used at a 1/500 dilution.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)
    Immunocytochemistry/ Immunofluorescence - Anti-c-Myc antibody [Y69] (ab32072)

    ICC/IF image of unpurified ab32072 stained HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32072, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

     

  • Flow Cytometry - Anti-c-Myc antibody [Y69] (ab32072)
    Flow Cytometry - Anti-c-Myc antibody [Y69] (ab32072)

    Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab32072 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32072, 1/76 dilution) for 30 min at 22ºC. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG [EPR25A] (monoclonal) (ab172730, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 nm bandpass filter. 

  • OI-RD Scanning - Anti-c-Myc antibody [Y69] (ab32072)
    OI-RD Scanning - Anti-c-Myc antibody [Y69] (ab32072)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-c-Myc antibody [Y69] (ab32072)
    Anti-c-Myc antibody [Y69] (ab32072)

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

数据表及文件

    • Datasheet
    • SDS
  • 文献 (602)

    发表研究结果有使用 ab32072?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab32072 被引用在 602 文献中.

    • Li C  et al. Homoharringtonine exhibits potent anti-tumor effect and modulates DNA epigenome in acute myeloid leukemia by targeting SP1/TET1/5hmC. Haematologica 105:148-160 (2020). PubMed: 30975912
    • Wang L  et al. A genetically defined disease model reveals that urothelial cells can initiate divergent bladder cancer phenotypes. Proc Natl Acad Sci U S A 117:563-572 (2020). PubMed: 31871155
    • Tang L  et al. Long Noncoding RNA DCST1-AS1 Promotes Cell Proliferation and Metastasis in Triple-negative Breast Cancer by Forming a Positive Regulatory Loop with miR-873-5p and MYC. J Cancer 11:311-323 (2020). PubMed: 31897227
    • Ballabio C  et al. Modeling medulloblastoma in vivo and with human cerebellar organoids. Nat Commun 11:583 (2020). PubMed: 31996670
    • Hu M  et al. Effects of electrical stimulation on cell activity, cell cycle, cell apoptosis and ß-catenin pathway in the injured dorsal root ganglion cell. Mol Med Rep 21:2385-2394 (2020). PubMed: 32323840
    View all Publications for this product

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    Western blot abreview for Anti-c-Myc antibody [Y69]

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (SK-N-AS)
    Loading amount
    30 µg
    Specification
    SK-N-AS
    Gel Running Conditions
    Reduced Denaturing (10% SDS gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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    提交于 May 03 2013

    Western blot abreview for Anti-c-Myc antibody [Y69]

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (multiple myeloma, MM.1S)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    20 µg
    Specification
    multiple myeloma, MM.1S
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    提交于 Mar 14 2019

    Immunoprecipitation abreview for Anti-c-Myc antibody [Y69]

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    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (HCT116 colon cancer cells)
    Total protein in input
    200 µg
    Treatment
    MG132
    Immuno-precipitation step
    Protein A
    Specification
    HCT116 colon cancer cells
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    提交于 Oct 02 2018

    Western blot abreview for Anti-c-Myc antibody [Y69]

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    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HCT116 colon cancer cells)
    Gel Running Conditions
    Reduced Denaturing (10)
    Loading amount
    20 µg
    Specification
    HCT116 colon cancer cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    提交于 Oct 02 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-c-Myc antibody [Y69]

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    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (brain tissue)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA ph 9
    Permeabilization
    No
    Specification
    brain tissue
    Blocking step
    Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
    Fixative
    Formaldehyde
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    Herr Dr. Markus Kipp

    Verified customer

    提交于 Aug 21 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-c-Myc antibody [Y69]

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    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Colorectal liver metastasis)
    Antigen retrieval step
    Heat mediated
    Permeabilization
    Yes - TBS with 0.25% Triton for 10 min
    Specification
    Colorectal liver metastasis
    Blocking step
    Power Block Universal Blocking Reagent 10x as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Formaldehyde
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    Mr. Michael Wuehrl

    Verified customer

    提交于 Jun 30 2017

    Immunohistochemistry (Frozen sections) abreview for Anti-c-Myc antibody [Y69]

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    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Dog Tissue sections (Colon)
    Permeabilization
    Yes - Triton
    Specification
    Colon
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
    Fixative
    Acetone
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    提交于 Apr 03 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-c-Myc antibody [Y69]

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    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (fetal liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: high pH CC1 (Roche Ventana)
    Specification
    fetal liver
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: 24°C
    Fixative
    Paraformaldehyde
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    提交于 Mar 16 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-c-Myc antibody [Y69]

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    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10 mM Citrate
    Sample
    Human Tissue sections (Colon)
    Specification
    Colon
    Permeabilization
    No
    Fixative
    Formaldehyde
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    提交于 May 30 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-c-Myc antibody [Y69]

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    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step
    None
    Sample
    Human Tissue sections (UBC)
    Specification
    UBC
    Permeabilization
    No
    Fixative
    Paraformaldehyde
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    提交于 Apr 02 2014

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