概述

  • 产品名称

    Anti-Myc tag抗体[9E10] - ChIP Grade
    参阅全部 Myc tag 一抗
  • 描述

    小鼠单克隆抗体[9E10] to Myc tag - ChIP Grade
  • 宿主

    Mouse
  • 特异性

    This antibody is specific for Myc tagged proteins. The Myc tag epitope (EQKLISEEDL) is located at the dimerization site of c-myc and therefore this antibody does not perform well at recognizing endogenous c-myc. A publication by Baker AM et al. 2016 (PubMed ID: 26826706 DOI: 10.1111/his.12939) shows the IHC staining generated by the 9E10 clone does not correlate with c-myc mRNA expression.

  • 经测试应用

    适用于: ICC/IF, ICC, ChIP, IHC (Methanol fixed), Flow Cyt, WB, IP, ELISA, IHC-P, IHC-Fr, Purificationmore details
  • 免疫原

    Synthetic peptide corresponding to Human Myc tag aa 400-500 (C terminal) conjugated to keyhole limpet haemocyanin.

  • 表位

    aa 410-419 of human Myc.
  • 阳性对照

    • Myc tagged proteins and myc tag expressing cells.
  • 常规说明

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

性能

应用

Our Abpromise guarantee covers the use of ab32 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
ICC/IF Use a concentration of 5 µg/ml.
ICC 1/2000.

See Abreviews.

ChIP Use at an assay dependent concentration.
IHC (Methanol fixed) 1/200. PubMed: 17329357
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB 1/500 - 1/1000.
IP Use at 6 µg/mg of lysate.
ELISA Use at an assay dependent concentration.
IHC-P 1/250 - 1/500.
IHC-Fr 1/1000. See Abreviews.
Purification Use at an assay dependent concentration.

靶标

  • 相关性

    Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein’s biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows anti-epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells.
  • 细胞定位

    Nuclear
  • 别名

    • c-myc tag antibody
    • Myc Epitope Tag antibody

图片

  • (A-D) Representative examples of body forming AtMORC7-MYC, AtMORC4-MYC, At-MORC1-MYC, and AtMORC6-MYC nuclei, respectively. (E) Untransformed wt nucleus subjected to the same antibody staining and imaging procedure. Left panels = anti-MYC channel; middle panels = DAPI channel (gray scaled). DAPI stains DNA, defining the position of dense chromocenters as high intensity white foci; right panels = merged channels (DAPI in blue, MYC in green). White triangles indicate examples of chromocenter adjacent AtMORC localization. Scale bars = 5 μM.

    Leaves from three-week old plants were fixed in 4% paraformaldehyde in TRIS buffer (10 mM TRIS pH 7.5, 10 mM EDTA, and 100 mM NaCl) for 20 minutes and washed twice in TRIS buffer. Leaves were chopped in 200–400 microliters lysis buffer (15 mM TRIS pH 7.5, 2 mM EDTA, 0.5 mM spermine, 80 mM KCl, 20 mM NaCl, and 0.1% Triton X-100) and filtered through a 3 μM cell strainer. 5 μL of nuclei suspension was added to 12 μL of sorting buffer (100mM TRIS pH 7.5, 50mM KCl, 2mM MgCl2, 0.05% Tween-20, and 20.5% sucrose) and air dried on chloroform dipped microscope slides for two hours and then post-fixed in 4% paraformaldehyde in PBS for 20 minutes. Slides were washed three times in PBS and incubated in blocking buffer (3% BSA, and 10% horse serum in PBS) for 30 minutes at 37°C. Nuclei were incubated at 4°C overnight in mouse monoclonal antibody against c-Myc (9E10, ab32; 1/200). Slides were washed in PBS and incubated with goat anti-mouse FITC antibody (ab7064; 1/200) for 90 minutes at room temperature. Following PBS washes, nuclei were counterstained and mounted in Vectashield mounting media with DAPI. Nuclei were analyzed with a Zeiss LSM 710 Confocal microscope at 63X or 100X magnification using Zen software.

     

  • Anti-Myc tag antibody [9E10] - ChIP Grade (ab32) at 1 µg/ml + E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) at 10 µg

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 45 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Lysate from E. coli recombinantly expressing 11 commonly used tags including myc tag.

  • Ab32 staining a Myc tagged protein in HeLa cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Endogenous c-myc was not detected under these conditions. Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton and blocked with 5% Serum for 30 minutes at 25°C. Samples were incubated with primary antibody (1/1000 in 5% Serum) for 1 hour at 25°C. An Alexa Fluor® 488 conjugated Goat anti-Mouse was used as a secondary antibody.

    See Abreview

  • RPE1 cells grown on coverslips were transfected with βarr2-myc, grown in low serum and then fixed and stained for Kif3A (red) and ab32 (green). Insets show higher magnifications of a representative PC. Kif3A was found in the cytoplasm and at the tip of the axoneme where it was colocalized with βarr2.

    Cells were incubated with primary antibodies in permeabilization buffer (PBS with 1 mg/mL bovine serum albumin (PBS-BSA) and 0.1% triton-X-100) for 45 minutes at room temperature. After two washes with PBS-BSA, cells were incubated for 30 minutes at room temperature in PBS-BSA containing secondary antibodies. After one wash with PBS-BSA and two washes in PBS, cells were laid down on microscope slides in a PBS–glycerol mix (50/50) with DAPI.

  • Phosphorylation of Cdc15 changes during the cell cycle. Exponentially growing cells (cyc) of CDC15-MYC9 (W1114) were arrested in G1 with a factor pheromone and released into fresh medium at 25°C. Cells were harvested at the indicated times, the percentage of divided nuclei was determined by DAPI staining of fixed cells, and proteins were analyzed by western blotting with 9E10 (ab32).

  • Cse4 histone variant occupancy at the endogenous CEN3 locus (eCEN3) and at a conditional CEN3 locus (cCEN3) when it is repressed (in the presence of galactose) or induced (glucose) by ChIP. Control (SC138) and Δfun30 (SC140) cells are grown in YP Gal until≈1 OD, then shifted in YP Glu or YP Gal containing 15 µg/ml of nocodazole and incubated for 4 hours. Cse4-Myc associated chromatin was immunoprecipitated and the immunoprecipitated DNA was analyzed by PCR followed by agarose gel electrophorsis and ethidium bromide staining to visualize the DNA fragments. Shown are the relevant, cropped out bands from a single, representative gel, input was 1/243th of the immunoprecipitated material.

    Overnight cultures grown in YPD at 30°C were diluted to 0.2 OD595, then grown to 0.7 OD595 at 30°C before crosslinking. Samples were crosslinked 15 min for H3, 3Myc-Htz1 and Cse4-Myc or 30 min for Fun30 detection with 1% final formaldehyde and chromatin extracts were sonicated to ∼500 bp. Triplicates or duplicate ChIP samples were validated by qPCR. Chromatin extracts were then immunoprecitated with 2 µg of mouse monoclonal anti-myc (9E10, ab32) for Cse4-Myc.

文献

This product has been referenced in:

  • Feng S  et al. Tripartite motif-containing 14 (TRIM14) promotes epithelial-mesenchymal transition via ZEB2 in glioblastoma cells. J Exp Clin Cancer Res 38:57 (2019). Read more (PubMed: 30728039) »
  • Crerar H  et al. Regulation of NGF Signaling by an Axonal Untranslated mRNA. Neuron 102:553-563.e8 (2019). Read more (PubMed: 30853298) »
See all 211 Publications for this product

客户评价及客户问答

21-30 of 44 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293 cells)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE gel)
Loading amount
10000 cells
Specification
HEK 293 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C

Mr. William Hung

Verified customer

提交于 Jul 26 2011

Application
Immunoprecipitation
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (W303 cells)
Total protein in input
100 µg
Immuno-precipitation step
Protein G
Specification
W303 cells

Abcam user community

Verified customer

提交于 Jul 26 2011

Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (BY4741 background)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE gel)
Loading amount
100000 cells
Specification
BY4741 background
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

提交于 Jun 17 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Esophagus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric buffer, pH6
Permeabilization
No
Specification
Esophagus
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Jun 02 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Saccharomyces cerevisiae Cell (BY4147 background)
Permeabilization
Yes - 0.25% Triton X-100 in PBS
Specification
BY4147 background
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Jun 01 2011

Application
ChIP
Sample
Saccharomyces cerevisiae Cell lysate - nuclear (YMV2 (genetically engineered strain to study DNA d)
Specification
YMV2 (genetically engineered strain to study DNA d
Detection step
Semiquantitative PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde

Abcam user community

Verified customer

提交于 May 31 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA - Buffer pH 9.0
Permeabilization
Yes - Wash Buffer Dako with Tween20
Specification
Tonsil
Fixative
Paraformaldehyde

Mr. Rudolf Jung

Verified customer

提交于 Mar 25 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (tonsil, cells near crypt)
Specification
tonsil, cells near crypt
Fixative
Formaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
1% BSA plus 5% normal serum in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

提交于 Apr 15 2009

Answer

Thank you for your enquiry. Ab32 recognizes recombinant proteins incorporating a c-Myc epitope tag and it also detects human c-Myc protein. The optimal dilution range to use in your samples does indeed depend on the abundance of protein in them; it is hard to tell what will work for your samples, I therefore recommend trying a range of dilutions in a preliminary experiment; I would recommend trying 1:100, 1:500, 1:1000, 1:2000 and optimise following the results of those tests.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (Hek 293 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Hek 293 cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

提交于 Aug 13 2007

21-30 of 44 Abreviews or Q&A

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