Anti-Radixin/RDX antibody [EP1862Y]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Radixin/RDX antibody [EP1862Y] (AB52495)

ICC/IF image of ab52495 stained Mcf7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52495, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Radixin/RDX antibody [EP1862Y] (AB52495)

Overlay histogram showing HeLa cells stained with ab52495 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52495, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• WB

Lab

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (AB52495)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (ab52495) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 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

Observed band size: 68 kDa

false

Exposure time: 59s

• WB

Lab

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (AB52495)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (ab52495) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 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

Observed band size: 68 kDa

false

Exposure time: 68s

• WB

Lab

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (AB52495)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (ab52495) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) transfected with scrambled siRNA control whole cell lysate at 10 µg

Lane 1:

RAW 264.7 transfected with siRNA specifically targeting RDX (Target name) whole cell lysate at 10 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 1/100000 dilution

Observed band size: 68 kDa

false

Exposure time: 8s

• WB

Unknown

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (AB52495)

All lanes:

Western blot - Anti-Radixin/RDX antibody [EP1862Y] (ab52495) at 1/1000 dilution

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

HRP labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 69 kDa

Observed band size: 80 kDa

false