Key features and details
- Rabbit monoclonal [EP1842Y] to FER
- Suitable for: WB, IP, Flow Cyt
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Product nameAnti-FER antibody [EP1842Y]
See all FER primary antibodies
DescriptionRabbit monoclonal [EP1842Y] to FER
SpecificityThis antibody reacts with FER
Tested applicationsSuitable for: WB, IP, Flow Cytmore details
Unsuitable for: ICC or IHC-P
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
Synthetic peptide within Human FER (N terminal). The exact sequence is proprietary.
- WB: Jurkat and HeLa cell lysate. Flow Cyt: Jurkat cells.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
KO cell lines
KO cell lysates
Our Abpromise guarantee covers the use of ab52479 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. Detects a band of approximately 93 kDa (predicted molecular weight: 93 kDa).|
|IP||Use a concentration of 5 µg/ml.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionTyrosine kinase of the non-receptor type. Probably performs an important function, perhaps in regulatory processes such as cell cycle control.
Tissue specificityExpressed in a variety of lymphoid cell lines.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. Fes/fps subfamily.
Contains 1 FCH domain.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Cellular localizationCytoplasm. Nucleus. Associated with the chromatin.
- Information by UniProt
- c FER antibody
- Feline encephalitis virus related kinase FER antibody
- Fer (fps/fes related) tyrosine kinase (phosphoprotein NCP94) antibody
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.
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.
All lanes : Anti-FER antibody [EP1842Y] (ab52479) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : FER knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 93 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Lanes 1 - 2: Merged signal (red and green). Green - ab52479 observed at 100 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab52479 was shown to react with FER in wild-type HeLa cells in Western blot with loss of signal observed in FER knockout cell line ab265226 (FER knockout cell lysate ab257950). Wild-type and FER knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab52479 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Anti-FER antibody [EP1842Y] (ab52479) at 1/5000 dilution + Jurkat cell lysate at 10 µg
Goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size: 93 kDa
Observed band size: 93 kDa
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
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab52479 has been referenced in 4 publications.
- 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 Lett 13:686-694 (2017). WB . PubMed: 28356947
- McGinnis LK et al. Fer tyrosine kinase is required for germinal vesicle breakdown and meiosis-I in mouse oocytes. Mol Reprod Dev 78:33-47 (2011). IHC-P, IF ; Mouse . PubMed: 21268181
- Whitehead SN et al. Transient and bilateral increase in Neuropilin-1, Fer kinase and collapsin response mediator proteins within membrane rafts following unilateral occlusion of the middle cerebral artery in mouse. Brain Res 1344:209-16 (2010). WB ; Mouse . PubMed: 20493826
- Fei F et al. The Fer tyrosine kinase regulates interactions of Rho GDP-Dissociation Inhibitor a with the small GTPase Rac. BMC Biochem 11:48 (2010). WB ; Human, Rat . PubMed: 21122136