- MA5-34806 - Invitrogen Antibodies RXRA antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Retinoid X Receptor alpha in MCF-7 cell. Samples were incubated with Retinoid X Receptor alpha monoclonal antibody (Product # MA5-34806), at a dilution of 1:5000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of RXR-alpha was performed by loading 100 µg of WT (lane 1) and RXR-alpha CRISPR KO (lane 2) HCT116 cell lysates in RIPA buffer onto a 4-15% gradient polyacrylamide gel. Proteins on the blots were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. RXR-alpha was detected at approximately 50 kDa using an RXR-alpha recombinant monoclonal antibody (Product # MA5-34806) at a dilution of 1:500 in 5% BSA in TBS with 0.1% Tween 20 (TBST) overnight at 4°C. The peroxidase-conjugated secondary antibody (Product # 65-6120) was diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Retinoid X Receptor alpha in HeLa cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with Retinoid X Receptor alpha monoclonal antibody (Product # MA5-34806), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Retinoid X Receptor alpha in A549 cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with Retinoid X Receptor alpha monoclonal antibody (Product # MA5-34806), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence of RXR-alpha was performed using HCT116 wild-type and RXR-alpha KO cells that were transfected with a green or a far-red fluorescent dye, respectively. Post-transfection, WT and KO cells were mixed and plated to a 1:1 ratio on coverslips as a mosaic and incubated for 24 hrs. Cells were fixed in 4% PFA (in PBS) or methanol for 15 min; cells were permeabilized with 0.1% Triton X-100 for 10 min at RT and blocked with PBS with 5% BSA, 5% goat serum, and 0.01% Triton X-100 for 30 min. Cells were stained with the RXR-alpha recombinant monoclonal antibody (Product # MA5-34806) at a 1:1,000 dilution overnight at 4°C. Secondary antibody incubation was performed using 1 µg/mL of Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 555 antibody (Product # A21429) together with DAPI for 1 hr. Imaging was performed with a 40X oil objective and analysis was performed using Image J. Cell image represents a single focal plane; WT and KO cells are outlined with a yellow (WT) or magenta (KO) dashed line. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Retinoid X Receptor alpha in paraffin-embedded rat liver tissue. Samples were incubated with Retinoid X Receptor alpha monoclonal antibody (Product # MA5-34806), and followed by hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Retinoid X Receptor alpha in paraffin-embedded human esophagus tissue. Samples were incubated with Retinoid X Receptor alpha monoclonal antibody (Product # MA5-34806), and followed by hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of RXR-alpha was performed on HCT116 cell lysates. Antibody-bead conjugates were prepared by adding 1 µg of RXR-alpha recombinant monoclonal antibody (Product # MA5-34806) with 30 µL of protein A-Sepharose beads and rocked overnight at 4°C. 1 mg of lysate was incubated with an antibody-bead conjugate for 2 hours at 4°C. Following centrifugation and multiple washes, 10% starting material (SM), 10% unbound fraction (UB) and immunoprecipitated fraction (IP) were processed for immunoblot using a different antibody (Product # 433900). Ponceau stained transfer of blot is shown. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

MA5-34806