Immunofluorescence analysis of EPO Receptor [pYpY454/456] was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ EPO Receptor [pYpY454/456] Recombinant Rabbit Oligoclonal Antibody (710484) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). F-actin (Panel b: red) was stained with Alexa Fluor® 594 Phalloidin (A12381). Nuclei (Panel c: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). Panel d is a merged image showing cytoplasmic and membrane localization. Panel e shows competition with the phospho- EPO R [pYpY454/456] peptide. The images were captured using Nikon enabled with cool snap at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Phosphorylation of the human EPO Receptor at the Tyr454 and Tyr456|
|Contains||0.09% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Receptor for erythropoietin. Mediates erythropoietin- induced erythroblast proliferation and differentiation. Upon EPO stimulation, EPOR dimerizes triggering the JAK2/STAT5 signaling cascade. In some cell types, can also activate STAT1 and STAT3. May also activate the LYN tyrosine kinase.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.