Anti-Rb antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rb antibody (AB226979)

Immunofluorescence analysis of HeLa cells fixed in 4% paraformaldehyde at RT for 15 min labelling Rb protein at nucleus using ab226979 at a 1/500 dilution (Green). phalloidin, a cytoskeleton marker, was stained at a 1/200 dilution (Red).
Scale bar= 10 µm.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb antibody (AB226979)

Paraffin-embedded human colon carcinoma tissue stained for Rb using ab226979 at 1/500 dilution in immunohistochemical analysis.

• IP

Supplier Data

Immunoprecipitation - Anti-Rb antibody (AB226979)

Rb was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with 4 μg of ab226979. Western blot was performed from the immunoprecipitate using ab226979 at 1/500 dilution.

Lane 1 : Rabbit IgG instead of ab226979 in HeLa whole cell lysate.

Lane 2 : ab226979 IP in HeLa whole cell lysate.

All lanes:

Immunoprecipitation - Anti-Rb antibody (ab226979)

Predicted band size: 106 kDa

false

• WB

Lab

Western blot - Anti-Rb antibody (AB226979)

Western blot : Anti-RB1 antibody (ab226979) staining at 1/1500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab226979 was shown to bind specifically to RB1. A band was observed at 120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in RB1 knockout cell line. To generate this image, wild-type and RB1 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Rb antibody (ab226979) at 1/1500 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lanes 1 - 4:

Western blot - Human RB1 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-rb1-knockout-hct116-cell-line-ab286657'>ab286657</a>)

Lane 2:

RB1 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type A549 ab288558 cell lysate at 20 µg

Lane 4:

RB1 knockout A549 <a href='/en-us/products/cell-lines/human-rb1-knockout-a549-cell-line-ab286470'>ab286470</a> cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 120 kDa

false

• WB

Lab

Western blot - Anti-Rb antibody (AB226979)

Western blot : Anti-RB1 antibody (ab226979) staining at 1/1500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab226979 was shown to bind specifically to RB1. A band was observed at 105 kDa in wild-type A549 cell lysates with no signal observed at this size in RB1 knockout cell line. To generate this image, wild-type and RB1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Rb antibody (ab226979) at 1/1500 dilution

Lane 1:

Wild-type A549 cell lysate at 40 µg

Lane 2:

RB1 knockout A549 cell lysate at 40 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

RB1 knockout HAP1 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

false

• WB

Supplier Data

Western blot - Anti-Rb antibody (AB226979)

Samples were separated by 5% SDS-PAGE.

All lanes:

Western blot - Anti-Rb antibody (ab226979) at 1/3000 dilution

All lanes:

Jurkat whole cell extract at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 106 kDa

false

• WB

Supplier Data

Western blot - Anti-Rb antibody (AB226979)

Samples were separated by 7.5% SDS-PAGE.

All lanes:

Western blot - Anti-Rb antibody (ab226979) at 1/3000 dilution

Lane 1:

Jurkat whole cell extracts at 30 µg

Lane 2:

Raji whole cell extracts at 30 µg

Lane 3:

NCI-H929 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 106 kDa

false