The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 98 kDa (predicted molecular weight: 84 kDa).
Use a concentration of 5 µg/ml.
Cellular uptake of iron occurs via receptor-mediated endocytosis of ligand-occupied transferrin receptor into specialized endosomes. Endosomal acidification leads to iron release. The apotransferrin-receptor complex is then recycled to the cell surface with a return to neutral pH and the concomitant loss of affinity of apotransferrin for its receptor. Transferrin receptor is necessary for development of erythrocytes and the nervous system (By similarity). A second ligand, the heditary hemochromatosis protein HFE, competes for binding with transferrin for an overlapping C-terminal binding site. Positively regulates T and B cell proliferation through iron uptake (PubMed:26642240). (Microbial infection) Acts as a receptor for new-world arenaviruses: Guanarito, Junin and Machupo virus.
Belongs to the peptidase M28 family. M28B subfamily. Contains 1 PA (protease associated) domain.
N- and O-glycosylated, phosphorylated and palmitoylated. The serum form is only glycosylated. Proteolytically cleaved on Arg-100 to produce the soluble serum form (sTfR). Palmitoylated on both Cys-62 and Cys-67. Cys-62 seems to be the major site of palmitoylation.
Secreted and Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Exposure time : 4 minutesTransferrin Receptor contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody (ab84036)Image courtesy of Carl Hobbs, King College London, U.K.
ab84036 immunohistochemical staining in the rat brain. Sampled were fixed with formaldehyade and heat-mediated antigen retrieval was performed with citric acid. Tissue sections were blocked in 1% BSA for 10 minutes at 21°C before incubation with ab84036 (1/6000) for 16 hours at 21°C. A biotin conjugated goat anti-rabbit IgG secondary was used at 1/250.
ICC/IF image of ab84036 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab84036 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - Anti-Transferrin Receptor antibody (ab84036)
Anti-Transferrin Receptor antibody (ab84036) at 1 µg/ml + Spleen (Mouse) Tissue Lysate at 10 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Exposure time : 8 minutesTransferrin Receptor contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Transferrin Receptor antibody (ab84036)This image is courtesy of an Abreview submitted by Ruma Raha-Chowdhury
ab84036 staining mouse liver sections by IHC-FrFl. The animal was perfused with 4% paraformaldehyde, further post fixed with 4% paraformaldehyde overnight and prepared for free floating sectioning. Staining with ab84036 at a 1/200 dilution in 0.1% TritonX with 0.1x PBS and 10% donkey was performed for 24h at 24°C. A donkey anti-rabbit Alexa568 polyclonal antibody was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Transferrin Receptor antibody (ab84036)Image courtesy of an anonymous Abreview.
ab84036 staining Transferrin Receptor in canine kidney cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde, permeabilized with 0.5% saponin then incubated with ab84036 at a 1/100 dilution for 1.5 hours at 20°C. The secondary used was an Alexa-Fluor 568 conjugated donkey anti-rabbit polyclonal, used at a 1/350 dilution.