Immunofluorescence analysis of PDGFR alpha was performed using 70% confluent log phase NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PDGFR alpha Rabbit Polyclonal Antibody (Product # PA5-16571) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing punctated cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide derived from the C-terminus of the precursor form of human PDGFR, alpha|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||1-3 µg/ml|
* 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.
PA5-16571 targets PDGFR alpha in IHC applications and shows reactivity with mouse, Rat, and Human samples.
The PA5-16571 immunogen is a synthetic peptide derived from the C-terminus of the precursor form of human PDGFR, alpha.
PDGFR (Platelet Derived Growth Factor Receptor) exhibits two different isoforms, alpha and beta; coded by two different genes. They possess five immunoglobulin like domains that are involved in ligand binding. Two tyrosine kinase domains are separated by a kinase insert to which PI-3 kinase can bind. PDGF AA can bind autophosphorylate and activate only PDGFR, alpha while PDGF BB can bind and activate to both PDGFR, alpha; and beta;.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.