- MA1-810A - Invitrogen Antibodies RARA antibody

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

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
Nucleic acids research 2021 Nov 18;49(20):11575-11595

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

Rogers MA, Chen J, Nallamshetty S, Pham T, Goto S, Muehlschlegel JD, Libby P, Aikawa M, Aikawa E, Plutzky J
Arteriosclerosis, thrombosis, and vascular biology 2020 Mar;40(3):656-669

Insoluble, speckled 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 2009 Jul;57(7):687-99

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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 4ºC 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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 ºC 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 ºC 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2. RAREs are required for RA signaling-induced Hoxa1 and Halr1 expression. ( A ) ChIP-seq data showing RARA and RXRA binding peaks at Halr1 and Hoxa1 loci in ESCs on day 1 of RA treatment. ( B ) JASPAR prediction of RAREs at the Halr1-Hoxa1 locus, the RARA/RXRA binding structural domain, and the CRISPR-Cas9-mediated RARE knockout scheme. ( C ) Agarose gel electrophoresis showing RARE knockout clones using primers shown in ( B ). ( D ) Sanger sequencing of PCR products in ( C ). ( E ) qRT-PCR showing Halr1 and Hoxa1 transcript levels in RARE knockout cells. Data represent the mean +- SD ( n = 4 replicates). Statistical significance was determined by a two-tailed Student's t -test (unpaired), * P < 0.05, ** P < 0.01. M: DNA ladder marker.

MA1-810A

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