Anti-ARID1B antibody [2D2]

• WB

Lab

Western blot - Anti-ARID1B antibody [2D2] (AB57461)

Western blot : Mouse Monoclonal[2D2] to ARID1B ab57461 staining at 1/1000 dilution, shown in green; Rabbit anti alpha Tubulin (ab52866) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 200 kDa in Wild-type HCT 116 Nuclear cell lysates with no signal observed at this size in ARID1B knockout HCT 116 Nuclear cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-ARID1B antibody [2D2] (ab57461) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 Nuclear at 20 µg

Lane 2:

Western blot - Human ARID1B knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-arid1b-knockout-hct116-cell-line-ab287219'>ab287219</a>) at 20 µg

Lane 3:

RKO at 20 µg

Lane 4:

PC-3 Membrane at 20 µg

Secondary

All lanes:

Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution

Predicted band size: 244 kDa

Observed band size: 200 kDa

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• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ARID1B antibody [2D2] (AB57461)

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labelling ARID1B (Green) with ab57461 at 10 µg/mL concentration.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ARID1B antibody [2D2] (AB57461)

Immunoperoxidase analysis of formalin-fixed paraffin-embedded human kidney tissue labelling ARID1B with ab57461 at 3 µg/mL concentration.

• IP

Supplier Data

Immunoprecipitation - Anti-ARID1B antibody [2D2] (AB57461)

Immunoprecipitation of ARID1B in HCT 116 cells. Lysates were prepared and immunoprecipitation was performed using 2 μg of ab57461 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

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:

Immunoprecipitation - Anti-ARID1B antibody [2D2] (ab57461) at 2 µg

All lanes:

HCT 116 cell lysates

Predicted band size: 244 kDa

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• WB

Supplier Data

Western blot - Anti-ARID1B antibody [2D2] (AB57461)

ab57461 was shown to react with ARID1B in wild-type HCT 116 cells in Western blot with loss of signal observed in ARID1B knockout cell line ab287219. Wild-type HCT 116 and ARID1B knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab57461 overnight at 4°C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/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-ARID1B antibody [2D2] (ab57461) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 lysate at 30 µg

Lane 2:

ARID1B knock-out HCT 116 lysate at 30 µg

Lane 2:

Western blot - Human ARID1B knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-arid1b-knockout-hct116-cell-line-ab287219'>ab287219</a>)

Observed band size: 244 kDa

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• WB

Supplier Data

Western blot - Anti-ARID1B antibody [2D2] (AB57461)

Blocking and diluting buffer and concentration : 5% non-fat milk/PBST (0.2%).

All lanes:

Western blot - Anti-ARID1B antibody [2D2] (ab57461) at 5 µg/mL

All lanes:

Western blot - Anti-ARID1B antibody [2D2] (ab57461) at 50 µg

Secondary

All lanes:

Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/5000 dilution

Predicted band size: 236 kDa

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• sELISA

Supplier Data

Sandwich ELISA - Anti-ARID1B antibody [2D2] (AB57461)

Sandwich ELSIA standard curve using ab57461 as a capture antibody. Detection limit for recombinant GST tagged ARID1B is 0.03 ng/mL.