Invitrogen

RARA Monoclonal Antibody (Ralpha10)

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RARA Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Retinoic Acid Receptor alpha (green) showing staining in the cytoplasm of NIH-3T3 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 fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

RARA Antibody in Western Blot (WB) — Western blot analysis of Retinoic Acid Receptor alpha was performed by loading 25 µg of SH-SY5Y (lane 1), MCF-7 (lane 2) and C6 (lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a Retinoic Acid Receptor alpha monoclonal antibody (Product # MA1-810A) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~55 kDa.

RARA Antibody in Immunohistochemistry (Frozen) (IHC (F)) — Published figure using RARA monoclonal antibody (Product # MA1-810A) in Immunohistochemistry (Frozen)

RARA Antibody in ChIP Assay (ChIP) — Figure 1. Hoxa1 and Halr1 are direct downstream target genes of RA signaling. ( A ) Schematic showing ESC differentiation protocols. ( B ) Images showing AP staining of undifferentiated and differentiated ESCs. Scale bars: 100 mum. ( C ) Bright-field images of EBs on days 1-6. Scale bars: 200 mum. ( D ) qRT-PCR data showing transcript levels of pluripotency and germ layer genes as well as Halr1 and Hoxa1 during ESC differentiation ( n = 3 replicates). ( E and F ) ChIP-seq data showing RARA and RXRA binding peaks at Halr1 and Hoxa1 loci on day 1 of RA-induced ESC differentiation. ( G ) qRT-PCR data showing Halr1 and Hoxa1 transcript levels in ESCs overexpressing RARA and RXRA on day 1 of RA treatment. Data represent the mean +- SD ( n = 3 replicates). Statistical significance was determined by a two-tailed Student's t -test (unpaired), * P < 0.05, ** P < 0.01. ( H ) qRT-PCR data showing Halr1 and Hoxa1 transcript levels in WT or KO on day 1 of RA treatment. Data represent the mean +- SD ( n = 3 replicates). Statistical significance was determined by a two-tailed Student's t -test (unpaired), * P < 0.05, ** P < 0.01.

Product Details

MA1-810A

Applications

Tested Dilution

Publications

Western Blot (WB)

1:100-1:1,000

Immunohistochemistry (Frozen) (IHC (F))

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

1:10-1:200

View 1 publication 1 publication

ChIP assay (ChIP)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Human, Rat

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

Ralpha10

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.

View immunogen

Conjugate

Unconjugated

Form

Lyophilized

Concentration

Conc. Not Determined

Storage buffer

ascites

Contains

0.05% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_11157462

Product Specific Information

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.

Target Information

RARA is a receptor for retinoic acid. Retinoic acid receptors bind as heterodimers to their target response elements in response to their ligands, all-trans or 9-cis retinoic acid, and regulate gene expression in various biological processes. The RXR/RAR heterodimers bind to the retinoic acid response elements (RARE) composed of tandem 5'-AGGTCA-3' sites known as DR1-DR5. In the absence of ligand, the RXR-RAR heterodimers associate with a multiprotein complex containing transcription corepressors that induce histone acetylation, chromatin condensation and transcriptional suppression. On ligand binding, the corepressors dissociate from the receptors and associate with the coactivators leading to transcriptional activation. RARA plays an essential role in the regulation of retinoic acid-induced germ cell development during spermatogenesis. Has a role in the survival of early spermatocytes at the beginning prophase of meiosis. In Sertoli cells, may promote the survival and development of early meiotic prophase spermatocytes. In concert with RARG, required for skeletal growth, matrix homeostasis and growth plate function. Regulates expression of target genes in a ligand-dependent manner by recruiting chromatin complexes containing KMT2E/MLL5. Mediates retinoic acid-induced granulopoiesis.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Enhancer architecture-dependent multilayered transcriptional regulation orchestrates RA signaling-induced early lineage differentiation of ESCs.

Nucleic acids research

Su G,Wang W,Zhao X,Chen J,Zheng J,Liu M,Bi J,Guo D,Chen B,Zhao Z,Shi J,Zhang L,Lu W

Published figure using RARA monoclonal antibody (Product # MA1-810A) in ChIP assay

Thu Nov 18 00:00:00 EST 2021

Retinoids Repress Human Cardiovascular Cell Calcification With Evidence for Distinct Selective Retinoid Modulator Effects.

Arteriosclerosis, thrombosis, and vascular biology

Rogers MA,Chen J,Nallamshetty S,Pham T,Goto S,Muehlschlegel JD,Libby P,Aikawa M,Aikawa E,Plutzky J

MA1-810A was used in Immunohistochemistry (Frozen) to investigate the roles of retinoic acid in osteogenic differentiation of human arterial smooth muscle cells and aortic valvular interstitial cells that participate in atherosclerosis and heart valve disease, respectively.

Sun Mar 01 00:00:00 EST 2020

Retinoids Repress Human Cardiovascular Cell Calcification With Evidence for Distinct Selective Retinoid Modulator Effects.

Arteriosclerosis, thrombosis, and vascular biology

Rogers MA,Chen J,Nallamshetty S,Pham T,Goto S,Muehlschlegel JD,Libby P,Aikawa M,Aikawa E,Plutzky J

Sun Mar 01 00:00:00 EST 2020

Insoluble, speckled cytosolic distribution of retinoic acid receptor alpha protein as a marker of hepatic stellate cell activation in vitro.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Mezaki Y,Yamaguchi N,Yoshikawa K,Miura M,Imai K,Itoh H,Senoo H

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

Wed Jul 01 00:00:00 EDT 2009

Bioinformatics

Protein Aliases: Nuclear receptor subfamily 1 group B member 1; nucleophosmin-retinoic acid receptor alpha fusion protein NPM-RAR long form; RAR Alpha; RAR alpha 1; RAR-alpha; retinoic acid; retinoic acid nuclear receptor alpha variant 1; retinoic acid nuclear receptor alpha variant 2; retinoic acid receptor; Retinoic acid receptor alpha; retinoic acid receptor, alpha polypeptide; RRA

Gene Aliases: NR1B1; RAR; RARA; RARalpha1

UniProt ID: (Human) P10276, (Mouse) P11416

Entrez Gene ID: (Human) 5914, (Rat) 24705, (Mouse) 19401

Molecular Function: C4 zinc finger nuclear receptor