Immunofluorescent analysis of Retinoic Acid Receptor alpha (green) showing staining in the cytoplasm of SH-SY5Y cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a Retinoic Acid Receptor alpha monoclonal antibody (Product # MA1-810A) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Synthetic peptide corresponding to residues C(444) S P S L S P S S H R S S P A T Q S P(462) of mouse RAR alpha.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:100-1:1000|
* 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.
|Immunocytochemistry (ICC)||See 1 publications below|
MA1-810A detects retinoic acid receptor (RAR) alpha from human, rat and mouse tissues and overexpressed cells. This antibody does not detect any of the RAR beta or gamma forms.
MA1-810A has been successfully used in Western blot and ICC/IF procedures. By Western blot, this antibody detects a single ~45 kDa protein representing RAR alpha in transfected COS cells. Due to the extremely low levels of RAR expressed in native tissues, it is recommended that sensitive detection systems, such as enhanced chemiluminescence, be used.
The apparent molecular weight of RAR alpha from transfected COS cells is ~45 kDa. In rat tissues, such as skin and testis, doublets in the ~50 kDa area are seen. RAR alpha has been identified with this antibody in human dermal and neonatal fibroblast cell lines, as well as mouse F9 and human SH-SY5Y cells; however, RAR alpha from human leukocytes and HUT102 cells were not identified by MA1-810 suggesting that there may be structural variation of RAR alpha between some cell types.
The MA1-810A immunogen is a synthetic peptide corresponding to residues C(444) S P S L S P S S H R S S P A T Q S P(462) of mouse RAR alpha. This peptide (Cat. # PEP-004) is available for use in neutralization and control experiments.
Reconstitute with PBS.
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 the retinoic acid receptor alpha is NR1B1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Insoluble, speckled cytosolic distribution of retinoic acid receptor alpha protein as a marker of hepatic stellate cell activation in vitro.
MA1-810A was used in immunocytochemistry to study the cytosolic distribution of retinoic acid receptor alpha protein as a marker of hepatic stellate cell activation in vitro
|Mezaki Y,Yamaguchi N,Yoshikawa K,Miura M,Imai K,Itoh H,Senoo H||The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (57:687)||2009|