Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] - BSA and Azide free (AB232472)

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (human breast adenocarcinoma epithelial cell) cells labeling Retinoid X Receptor alpha/RXRA with Purified ab125001 at 1 : 500 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG (Alexa Fluor^® 488) was used as the secondary antibody at 1 : 1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125001).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] - BSA and Azide free (AB232472)

This data was developed using the same antibody clone in a different buffer formulation (ab125001). ab125001 staining Retinoid X Receptor alpha in wild-type HCT116 cells (top panel) and RXRA knockout HCT116 cells (bottom panel) (ab273708). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab125001 at 0.2μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• IP

Lab

Immunoprecipitation - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] - BSA and Azide free (AB232472)

ab125001 (purified) at 1 : 20 dilution (0.6μg) immunoprecipitating Retinoid X Receptor alpha/RXRA in HeLa whole cell lysate.

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10μg.
Lane 2 (+) : ab125001 & HeLa whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab125001 in HeLa whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125001).

All lanes:

Immunoprecipitation - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (<a href='/en-us/products/primary-antibodies/retinoid-x-receptor-alpha-rxra-antibody-epr7106-ab125001'>ab125001</a>)

Predicted band size: 51 kDa

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• WB

Lab

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] - BSA and Azide free (AB232472)

This data was developed using the same antibody clone in a different buffer formulation (ab125001)

ab125001 was shown to react with RXRA in wild-type HCT116 cells in Western blot with loss of signal observed in RXRA knockout cell line ab273708. Wild-type HCT116 and RXRA knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab125001 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2ug/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (<a href='/en-us/products/primary-antibodies/retinoid-x-receptor-alpha-rxra-antibody-epr7106-ab125001'>ab125001</a>) at 1/500 dilution

Lane 1:

Wild-type HCT116 lysate at 20 µg

Lane 2:

Western blot - Human RXRA knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-rxra-knockout-hct116-cell-line-ab273708'>ab273708</a>) at 20 µg

Observed band size: 51 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] - BSA and Azide free (AB232472)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.