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  1. Link

    products/primary-antibodies/myc-tag-antibody-9e10-ab32.pdf

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Tags & Cell Markers Epitope Tags Myc Tag
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Anti-Myc tag antibody [9E10] (ab32)

  • Datasheet
  • SDS
Reviews (29)Q&A (17)References (338)

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Western blot - Anti-Myc tag antibody [9E10] (ab32)
  • Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)
  • Western blot - Anti-Myc tag antibody [9E10] (ab32)
  • Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)
  • Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)
  • Western blot - Anti-Myc tag antibody [9E10] (ab32)

Key features and details

  • Mouse monoclonal [9E10] to Myc tag
  • Suitable for: ICC/IF, Flow Cyt, WB, IP, ELISA, IHC-Fr, Purification
  • Reacts with: Recombinant fragment
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 APC Biotin Biotin DyLight® 488 DyLight® 550 DyLight® 650 FITC HRP PE SureLight® APC

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Overview

  • Product name

    Anti-Myc tag antibody [9E10]
    See all Myc tag primary antibodies
  • Description

    Mouse monoclonal [9E10] to Myc tag
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WB, IP, ELISA, IHC-Fr, Purificationmore details
  • Species reactivity

    Reacts with: Recombinant fragment
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Myc tagged proteins and myc tag expressing cells.
  • General notes

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    9E10
  • Myeloma

    Sp2/0
  • Isotype

    IgG1
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Epitope Tags
    • Myc Tag
    • 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

Associated products

  • Alternative Versions

    • DyLight® 650 Anti-Myc tag antibody [9E10] (ab117487)
    • DyLight® 488 Anti-Myc tag antibody [9E10] (ab117499)
    • DyLight® 550 Anti-Myc tag antibody [9E10] (ab117511)
    • FITC Anti-Myc tag antibody [9E10] (ab117599)
    • Biotin Anti-Myc tag antibody [9E10] - C-terminal (ab197139)
    • Alexa Fluor® 488 Anti-Myc tag antibody [9E10] (ab202008)
    • Anti-Myc tag antibody [9E10] (ab206486)
    • Alexa Fluor® 594 Anti-Myc tag antibody [9E10] (ab223894)
    • Alexa Fluor® 647 Anti-Myc tag antibody [9E10] (ab223895)
    • APC Anti-Myc tag antibody [9E10] (ab223896)
    • Anti-Myc tag antibody [9E10] - BSA and Azide free (ab264603)
    • HRP Anti-Myc tag antibody [9E10] (ab62928)
    • PE Anti-Myc tag antibody [9E10] (ab72468)
    • SureLight® APC Anti-Myc tag antibody [9E10] (ab72580)
    • Biotin Anti-Myc tag antibody [9E10] (ab81658)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
ICC/IF (6)
Use a concentration of 5 µg/ml.
Flow Cyt
Use 1µg for 106 cells.

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

WB (13)
1/500 - 1/1000.
IP (3)
Use at 6 µg/mg of lysate.
ELISA
Use at an assay dependent concentration.
IHC-Fr
1/1000.

See Abreviews.

Purification
Use at an assay dependent concentration.
Notes
ICC/IF
Use a concentration of 5 µg/ml.
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-Fr
1/1000.

See Abreviews.

Purification
Use at an assay dependent concentration.

Target

  • Relevance

    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.
  • Cellular localization

    Nuclear
  • Alternative names

    • c-myc tag antibody
    • Myc Epitope Tag antibody

Images

  • Western blot - Anti-Myc tag antibody [9E10] (ab32)
    Western blot - Anti-Myc tag antibody [9E10] (ab32)
    All lanes : Anti-Myc tag antibody [9E10] (ab32) at 1/200 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : MYC CRISPR-Cas9 edited HEK-293T cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : SH-SY5Y cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.


    False colour image of Western blot: Anti-Myc tag antibody [9E10] staining at 1/200 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32 was shown to bind specifically to Myc tag. A band was observed at 57 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in MYC CRISPR-Cas9 edited cell line ab256500 (CRISPR-Cas9 edited cell lysate ab263850). The band observed in the CRISPR-Cas9 edited lysate lane below 57 kDa is likely to represent a truncated form of Myc tag. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MYC CRISPR-Cas9 edited HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)
    Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)Image from Harris CJ et al., PLoS Genet. 2016;12(5):e1005998. Fig 5.; doi: 10.1371/journal.pgen.1005998. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    (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.

     

  • Western blot - Anti-Myc tag antibody [9E10] (ab32)
    Western blot - Anti-Myc tag antibody [9E10] (ab32)
    Anti-Myc tag antibody [9E10] (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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)
    Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)This image is courtesy of an anonymous Abreview

    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

  • Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)
    Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody [9E10] (ab32)Image from Molla-Herman A et al., PLoS One. 2008;3(11):e3728. Fig 8(B).; doi: 10.1371/journal.pone.0003728. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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.

  • Western blot - Anti-Myc tag antibody [9E10] (ab32)
    Western blot - Anti-Myc tag antibody [9E10] (ab32)Menssen R et al., (2001) Curr Biol. Mar 6;11(5):345-50.

    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).

Protocols

  • Western Blot
  • Mouse on Mouse staining protocol

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (338)

Publishing research using ab32? Please let us know so that we can cite the reference in this datasheet.

ab32 has been referenced in 338 publications.

  • Forey R  et al. A Role for the Mre11-Rad50-Xrs2 Complex in Gene Expression and Chromosome Organization. Mol Cell 81:183-197.e6 (2021). PubMed: 33278361
  • Akkouche A  et al. In vivo antagonistic role of the Human T-Cell Leukemia Virus Type 1 regulatory proteins Tax and HBZ. PLoS Pathog 17:e1009219 (2021). PubMed: 33471856
  • Zhou X  et al. Cross-compartment signal propagation in the mitotic exit network. Elife 10:N/A (2021). PubMed: 33481703
  • Guo ZF & Kong FL Akt regulates RSK2 to alter phosphorylation level of H2A.X in breast cancer. Oncol Lett 21:187 (2021). PubMed: 33574926
  • Zhou X  et al. NCK1-AS1 promotes the progression of lung squamous cell carcinoma through transcriptionally upregulating NCK1 via interacting with MYC. Cancer Biol Ther 22:196-203 (2021). PubMed: 33629937
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-10 of 46 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Myc tag antibody [9E10]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RM-1 - mouse prostate cancer cells)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
RM-1 - mouse prostate cancer cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Dec 03 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Myc tag antibody [9E10]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
HeLa
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 06 2020

ChIP abreview for Anti-Myc tag antibody [9E10] - ChIP Grade

Good
Abreviews
Abreviews
Application
ChIP
Sample
Human Cell lysate - whole cell (HEK cells)
Specification
HEK cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 12 2017

Mass Cytometry abreview for Anti-Myc tag antibody [9E10]

Excellent
Abreviews
Abreviews
Application
Mass Cytometry
Sample
Mouse Cell (Lung)
Specification
Lung
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Irit Milman Krentsis

Verified customer

Submitted Nov 14 2017

Western blot abreview for Anti-Myc tag antibody [9E10] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
30 µg
Specification
HEK cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 13 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Myc tag antibody [9E10] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton
Specification
HeLa
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 05 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Myc tag antibody [9E10] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (Kidney)
Permeabilization
Yes - 0.1% Tween-20
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Thomas Kidd

Verified customer

Submitted Jul 14 2017

Western blot abreview for Anti-Myc tag antibody [9E10] - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 06 2016

Western blot abreview for Anti-Myc tag antibody [9E10] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (LNCaP)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
40 µg
Specification
LNCaP
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 09 2015

Western blot abreview for Anti-Myc tag antibody [9E10] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Primary lung cancer line LKR10)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
Primary lung cancer line LKR10
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 25 2014

1-10 of 46 Abreviews or Q&A

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