Immunofluorescence analysis of Retinoic Acid Receptor alpha was performed using 70% confluent SH-SY5Y cells treated with 10µM retinoic acid for 72 hours. 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 RARA Rabbit Polyclonal Antibody (PA1-810A) at 1:250 dilution 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 conjugate (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 (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e is untreated cell with no signal. Panel f represents control cells with no primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C S(444) P S L S P S S H R S S P A T Q S P (462) of mouse RAR alpha.|
|Storage buffer||whole serum diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1,000|
* 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.
PA1-810A detects retinoic acid receptor (RAR) from human and mouse tissues and cells. This antibody does not detect RAR beta or gamma forms.
PA1-810A has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~50-58 kDa protein representing RAR alpha from Jurkat, CEM-C7, and IM9 cells.
The PA1-810A immunogen is a synthetic peptide corresponding to residues C S(444) P S L S P S S H R S S P A T Q S P (462) of mouse RAR alpha. PA1-810A immunizing peptide (Cat. # PEP-004) is available for use in neutralization and control experiments.
Retinoic acid, a metabolite of vitamin A, is necessary for normal organogenesis but acts as a teratogen at high levels during embryonic and fetal development. Retinoic acid functions through its interaction with the nuclear protein, retinoic acid receptor (RAR). RAR belongs to the steroid and thyroid hormone superfamily of nuclear receptor proteins which exert their effects by binding to specific DNA response elements, thus regulating gene expression in target cells. RAR exists as three major subtypes: alpha, beta and gamma. The corresponding gene for the retinoic acid receptor alpha is NR1B1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.