Anti-Myc tag antibody [9E10] - ChIP Grade (ab32)

Overview

  • Product name
    Anti-Myc tag antibody [9E10] - ChIP Grade
    See all Myc tag primary antibodies
  • Description
    Mouse monoclonal [9E10] to Myc tag - ChIP Grade
  • Specificity
    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.
  • Tested applications
    Suitable for: ICC/IF, ICC, ChIP, IHC (Methanol fixed), Flow Cyt, WB, IP, ELISA, IHC-P, IHC-Fr, Purificationmore details
  • Immunogen

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

  • Epitope
    aa 410-419 of human Myc.
  • Positive control
    • Myc tagged proteins and myc tag expressing cells.

Properties

Applications

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 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.

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

  • 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.

    Predicted band size : 41 kDa
    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



  • Predicted band size : 41 kDa

    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).

References

This product has been referenced in:
  • Jiang H  et al. The regulator of calcineurin 1 increases adenine nucleotide translocator 1 and leads to mitochondrial dysfunctions. J Neurochem 140:307-319 (2017). WB ; Human . Read more (PubMed: 27861892) »
  • Cheung RS  et al. Ubiquitination-Linked Phosphorylation of the FANCI S/TQ Cluster Contributes to Activation of the Fanconi Anemia I/D2 Complex. Cell Rep 19:2432-2440 (2017). WB ; Human . Read more (PubMed: 28636932) »

See all 132 Publications for this product

Customer reviews and Q&As

Application
CyTOF
Sample
Mouse Cell (Lung)
Specification
Lung
Username

Irit Milman

Verified customer

Submitted Nov 14 2017

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
Username

Abcam user community

Verified customer

Submitted Nov 13 2017

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
Username

Abcam user community

Verified customer

Submitted Sep 05 2017

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
Username

Dr. T Kidd

Verified customer

Submitted Jul 14 2017

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
Username

Abcam user community

Verified customer

Submitted Jul 06 2016

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
Username

Abcam user community

Verified customer

Submitted Jan 09 2015

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
Username

Abcam user community

Verified customer

Submitted Jul 25 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (Lung cancer line A549)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
Lung cancer line A549
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 25 2014

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

Abcam user community

Verified customer

Submitted Jul 15 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing (6%)
Loading amount
30 µg
Specification
HeLa
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jul 04 2013

1-10 of 43 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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