ELISA: Antibody specificity was verified by ELISA against the peptide (EQKLISEEDL). A 1:60,000 dilution of the antibody gave an O.D.=1.0 in a 15 minute reaction using HRP-conjugated Goat Anti Rabbit IgG at 1:20,000 and TMB as the substrate. Appropriate specificity controls were run.
Use at an assay dependent concentration. PubMed: 19793804
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration. PubMed: 16093346
Use at an assay dependent concentration.
Use at an assay dependent concentration. See Abreview for further details (submitted by Eeva-Liisa Eskelinen).
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.
c-myc tag antibody
Myc Epitope Tag antibody
Immunocytochemistry/ Immunofluorescence - Anti-Myc tag antibody (ab9106)This image is courtesy of an anonymous Abreview
ab9106 staining the Myc tag in HeLa cells transfected with a Myc-tagged membrane targeting protein by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X (0.5%) and blocked with 3% BSA for 1 hour at 24°C. Samples were incubated with primary antibody (1/500) for 1 hour at 24°C. An Alexa Fluor®488-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.
Immunoprecipitation - Anti-Myc tag antibody (ab9106)This image is courtesy of an anonymous abreview.
ab9106 at 4µg/mg lysate used in human huh-7 whole cell lysate (1x106 cells). Cells expressed myc-tagged CDC42 protein. Immunoprecipitation step performed using Protein A matrix. Incubation time 4 hours at 4°C. Western Blot antibody used at 1/300 dilution.
Immunoprecipitation - Anti-Myc tag antibody (ab9106)
Ab9106 used at 1/250 in the immunoprecipitation of transfected human 293FT cells (whole cell lysate). In this experiment cells were co-transfected with myc tagged cdc25 along with an HA tagged viral protein which binds to cdc25. The cell lysate was immunoprecipitated using ab9106, run on a 10% gel. The blot was probed for HA and HA tagged protein (which co-IPed with Myc tagged protein) and this is seen at ~15KDa.
Lane 1: Input untransfected cells
Lane 2: Immunoprecipitated untransfected cells
Lane 3: Input co-transfected cells
Lane 4: Immunoprecipitated co-transfected cells
This image is courtesy of an Abreview submitted on 23 March 2006.
Electron Microscopy - Anti-Myc tag antibody (ab9106)This image is courtesy of an Abreview submitted by Dr Eeva-Liisa Eskelinen
ab9106 at 1/2000 staining mouse embryonic fibroblasts expressing myc-tagged protein by ICC. The cells were paraformaldehyde fixed and blocked with 3% BSA prior to incubation with the antibody for 1 hour. A goat anti-rabbit IgG antibody (10nm Gold conjugated) was used as the secondary. The image shown is immuno electron microscopical staining with thawed cryosections.