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
  • 特异性This antibody is specific for endogenous c-Myc.
  • 经测试应用适用于: WB, ICC/IF, Flow Cyt, IHC-P, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

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

  • 阳性对照
    • 购买相配的WB阳性对照Recombinant human c-Myc protein
    • WB: Jurkat, Raji, K562, THP1, A20 and Raw264.7 cell lysates. ICC/IF: HeLa cells. IHC-P: Human skin carcinoma, diffuse large B cell lymphoma, adenocarcinoma of the colon, lung adenocarcinoma, gastric adenocarcinoma, urinary bladder transitional carcinoma tissues and esophagus. IP: Jurkat cell lysate. Flow Cyt: HeLa cells.
  • 常规说明

    Myc is involved in MAPK-p38 signaling pathway - see the interactive version. If you need conjugated anti-c-myc (Y69) RabMAb antibodies, find our range of products here.

    We also offer a PBS only version of this clone as product ab168727.

     

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

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.

应用 Ab评论 说明
WB 1/10000. Detects a band of approximately 57 kDa (predicted molecular weight: 49 kDa).Can be blocked with c-Myc peptide (ab166837).
ICC/IF Use a concentration of 10 µg/ml.

1/100.

Flow Cyt 1/76.
IHC-P Use a concentration of 5 µg/ml.

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.
  • Information by UniProt
  • 数据库链接
  • 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
    • Class E basic helix-loop-helix protein 39 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
    • MYCC antibody
    • Myelocytomatosis oncogene antibody
    • Niard antibody
    • Nird antibody
    • Oncogene Myc antibody
    • OTTHUMP00000158589 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
    see all

Anti-c-Myc antibody [Y69] 图像

  • 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.

  • ab32072 staining c-Myc in HeLa 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).

  • Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling c-Myc with ab32072 at 1/500 dilution,followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.Nuclear staining on mouse spleen.Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • 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.

  • Overlay histogram showing HeLa 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 Alexa Fluor® 488 goat anti-rabbit IgG (H&L) preadsorbed (ab150081) 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. 

  • Immunocytochemistry/immunofluorescence analysis of HeLa cells labelling c-Myc with purified ab32072 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

  • 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.

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  • ICC/IF image of unpurified ab32072 stained HeLa 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 anti rabbit DyLight® 488 IgG - H&L, pre-adsorbed (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.

  • All lanes : Anti-c-Myc antibody [Y69] (ab32072) at 1/10000 dilution (purified)

    Lane 1 : Raji cell lysate
    Lane 2 : K562 cell lysate
    Lane 3 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • 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
    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.

  • 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. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) 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) 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. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) 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) analysis of human skin carcinoma tissue labelling c-Myc with unpurified ab32072 at 1/50.

  • 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) analysis of human lung adenocarinoma tissue labelling c-Myc with unpurified ab32072.

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

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human urinary bladder transitional carcinoma tissue labelling c-Myc with unpurified ab32072.

  • c-Myc was immunoprecipitated using 0.5mg Jurkat 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]

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

Anti-c-Myc antibody [Y69] (ab32072)参考文献

This product has been referenced in:
  • Franco M  et al. A Novel Secreted Protein, MYR1, Is Central to Toxoplasma's Manipulation of Host Cells. MBio 7:e02231-15 (2016). WB . Read more (PubMed: 26838724) »
  • Agarwal R  et al. Role of immunohistochemistry in the era of genetic testing in MYC-positive aggressive B-cell lymphomas: a study of 209 cases. J Clin Pathol 69:266-70 (2016). Read more (PubMed: 26307073) »

See all 129 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 May 03 2013

The antibody ab32072 would be to detect the c-myc protein.

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
Username

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提交于 May 30 2014

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
Username

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Verified customer

提交于 Apr 02 2014

Application Western blot
Loading amount 15 µg
Gel Running Conditions Reduced Denaturing (any kDa gel)
Sample Human Cell lysate - whole cell (A431 cells)
Specification A431 cells
Treatment EGF for 24 hours
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Felicia Cooper

Verified customer

提交于 Mar 19 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Pancreas)
Specification Pancreas
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate pH 6
Permeabilization No
Blocking step 1% BSA 1% FBS as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

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提交于 Mar 13 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (SW480 cells, HCT116 cells)
Loading amount 100000 cells
Specification SW480 cells, HCT116 cells
Gel Running Conditions Reduced Denaturing (4-12%)
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 Dec 17 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (BJAB)
Total protein in input 10 µg
Immuno-precipitation step Protein A/G
Specification BJAB
Username

Dr. Meztli Arguello

Verified customer

提交于 Jul 11 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Lymphocytes)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 14 µg
Specification Lymphocytes
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 May 12 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (Canine B cell lymphoma)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification Canine B cell lymphoma
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
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提交于 Dec 23 2015

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