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Anti-FER antibody [EP1842Y] images
Western blot - Anti-FER antibody [EP1842Y] (ab52479)
Predicted band size : 93 kDa
Lane 1: Wild-type HAP1 cell lysate (40 µg) Lane 2: FER knockout HAP1 cell lysate (40 µg) Lane 3: HeLa cell lysate (20 µg) Lane 4: Jurkat cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab52479 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab52479 was shown to specifically react with FER when FER knockout samples were used. Wild-type and FER knockout samples were subjected to SDS-PAGE. Ab52479 and ab8245 (loading control to GAPDH) were diluted at 1/5000 and 1:10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1:10,000 dilution for 1 hour at room temperature before imaging.
Western blot - FER antibody [EP1842Y] (ab52479)
Anti-FER antibody [EP1842Y] (ab52479) at 1/5000 dilution + Jurkat cell lysate at 10 µg
Secondary Goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size : 93 kDa Observed band size : 93 kDa
Overlay histogram showing Jurkat cells stained with ab52479 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52479, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
FER was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to FER and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52479. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Band: 93kDa; FER
References for Anti-FER antibody [EP1842Y] (ab52479)
This product has been referenced in:
Hu X et al. Downregulation of feline sarcoma-related protein inhibits cell migration, invasion and epithelial-mesenchymal transition via the ERK/AP-1 pathway in bladder urothelial cell carcinoma. Oncol Lett13:686-694 (2017).
Read more (PubMed: 28356947) »
McGinnis LK et al. Fer tyrosine kinase is required for germinal vesicle breakdown and meiosis-I in mouse oocytes. Mol Reprod Dev78:33-47 (2011).
Read more (PubMed: 21268181) »