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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Src.
(Peptide available as
This product is a recombinant rabbit monoclonal antibody.
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.
A trial size is available to purchase for this antibody.
Our Abpromise guarantee covers the use of ab32078 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000 - 1/10000. Predicted molecular weight: 60 kDa.Can be blocked with Src (phospho Y529) peptide (ab179556).|
|ICC/IF||1/50 - 1/500.|
WB analysis. Lane 1:Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates 15ug and Lane 2:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H2O2 at 10mM for 1 h. whole cell lysates 15ug and Lane 3:HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with H2O2 at 10mM for 1 h. whole cell lysates 15ug. Then the membrane was incubated with phosphatase. Primary ab used at 1:30,000 dilution. 5% NFDM/TBST was used as Blocking buffer and Diluting buffer. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as a Secondary ab at 1:20,000 dilution. The Exposure time was 30 seconds.
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling Src (phospho Y529) with ab32078 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200) - Microtubule Marker (Alexa Fluor® 594). DAPI (blue) was used as a nuclear counterstain.
The green staining was increased in the H2O2 (10mM, 1 hour) treated HeLa cells when compared with HeLa cells without treatment. After LP treatment, the green signaling was obviously decreased.
For the pan antibody, there was no great difference after H2O2 (10 mM, 1 hour) or EGF (100 ng/mL, 10 minutes) + LP treatment.
The data showed mostly Cytoplasm and Membran staining for Ab32078.
Dot Blot analysis of Lane 1: Src (pY529) phospho peptide and Lane 2: Src non-phospho peptide labeling Src (phospho Y529) with ab32078 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.