Human ARID1B knockout HCT116 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Supplier Data
Western blot - Human ARID1B knockout HCT116 cell line (AB287219)
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] (<a href='/en-us/products/primary-antibodies/arid1b-antibody-2d2-ab57461'>ab57461</a>) 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 (ab287219)
Observed band size: 244 kDa
false
- WB
Lab
Western blot - Human ARID1B knockout HCT116 cell line (AB287219)
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] (<a href='/en-us/products/primary-antibodies/arid1b-antibody-2d2-ab57461'>ab57461</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 Nuclear at 20 µg
Lane 2:
Western blot - Human ARID1B knockout HCT116 cell line (ab287219) 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
false
- NGS
Lab
Next Generation Sequencing - Human ARID1B knockout HCT116 cell line (AB287219)
1 bp insertion and 92 bp deletion (allele 1), 50 bp deletion (allele 2) in exon 11 of CCDS5251.2
Complementary Products
Cell Lines & Lysates
AB266889
Human POR (Cytochrome P450 Reductase) knockout HCT116 cell line
Advanced Validation
- 0
- 0
Primary Antibodies
AB274322
Anti-htrA1 antibody [EPR23240-64]
20ul selling size|KO Validated|RabMAb|Recombinant|Trial Size
![Immunoprecipitation - Anti-htrA1 antibody [EPR23240-64] (AB274322)](https://content.abcam.com/products/images/htra1-antibody-epr23240-64-ab274322--immunoprecipitation-img8296.jpg)
- 0
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Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
The protein participates in the regulation of transcription by altering the chromatin architecture. As part of the SWI/SNF complex ARID1B works alongside other proteins to control which parts of the genome are accessible for transcription. Its activity influences numerous cellular events like cell differentiation and proliferation. The SWI/SNF complex plays a significant role in embryonic development and tumor suppression.
Pathways
ARID1B affects several critical biological processes such as Wnt signaling and cell cycle regulation. The protein interacts with beta-catenin in the Wnt pathway affecting gene expression and cell fate decisions. Additionally ARID1B modulates the cell cycle directly by influencing the expression of cyclins and cyclin-dependent kinases proteins essential for cell cycle progression.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.<br>3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
McCoY5a + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com