Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (AB125001)

Immunocytochemistry 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (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] (AB125001)

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.

All lanes:

Immunoprecipitation - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (ab125001)

Predicted band size: 51 kDa

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• WB

Lab

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (AB125001)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (ab125001) at 1/1000 dilution

Lane 1:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 16 kDa,26 kDa,51 kDa

Observed band size: 14 kDa,35 kDa

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• WB

Lab

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (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] (ab125001) 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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• WB

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Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (AB125001)

Lanes 1 - 4 : Merged signal (red and green). Green - ab125001 observed at 53 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab125001 was shown to react with Anti-Retinoid X Receptor alpha in wild-type HCT 116 cells in western blot with loss of signal observed in RXRA knockout cell line ab273708 (RXRA knockout cell lysate ab275245). Wild-type and RXRA knockout HCT 116 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab125001 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (ab125001) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

RXRA knockout HCT116 cell 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>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 51 kDa

Observed band size: 53 kDa

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• WB

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Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (AB125001)

All lanes:

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (ab125001) at 1/1000 dilution

Lane 1:

MCF7 cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

K562 cell lysate at 10 µg

Lane 4:

RAW 264.7 cell lysate at 10 µg

Lane 5:

PC12 cell lysate at 10 µg

Lane 6:

NIH 3T3 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted 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] (AB125001)

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.

• WB

CiteAb

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (AB125001)

Retinoid X Receptor alpha/RXRA western blot using anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] ab125001. Publication image and figure legend from Zhang, S., Jin, T., et al., 2020, Front Cell Neurosci, PubMed 32792909.

ab125001 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab125001 please see the product overview.

Electro-acupuncture promotes the release of exosomal miR-146b in peri-ischemic striatum. (A) The expression of exosome-associated markers HSP70, TSG101, and CD81 were analyzed by Western blot. (B) The expression of markers were quantified on the basis of average band intensity (n = 4). (C) MiRNA microarray analysis of exosomes from peri-ischemic striatum and clustering of 25 significantly altered miRNAs with ≥1.5-fold difference in expression between MCAO and MCAO+EA groups. (D) Four miRNAs were significantly upregulated and 1 miRNA were downregulated in exosomes of the MCAO+EA group compared with the MCAO group were checked by RT-qPCR (n = 3). (E) The expression of miR-146b in tissues of peri-ischemic striatum were analyzed form MCAO, MCAO+miR-146b inhibitors, MCAO+EA, MCAO+EA+miR-146b inhibitors groups (n = 3). Data represent means ± SD of the results of representative experiment. *P< 0.05, MCAO vs. Sham; **p < 0.01, MCAO vs. Sham; #P< 0.05 and ##P< 0.01, MCAO vs. MCAO+EA; &&P< 0.01, MCAO vs. MCAO+miR-146b inhibitors; MCAO+EA vs. MCAO+EA+miR-146b inhibitors.

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