Human ARNTL knockout MCF7 cell line
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- WB
Lab
Western blot - Human ARNTL knockout MCF7 cell line (AB289293)
Western blot : Rabbit monoclonal [EPR23696-22] to BMAL1 ab230822 staining at 1/500 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.
A band was observed at 69-75 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in ARNTL knockout MCF7 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-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-BMAL1 antibody [EPR23696-22] (<a href='/en-us/products/primary-antibodies/bmal1-antibody-epr23696-22-ab230822'>ab230822</a>) at 1/500 dilution
Lane 1:
Wild-type MCF7 at 20 µg
Lane 2:
ARNTL knockout MCF7 at 20 µg
Lane 2:
Western blot - Human ARNTL knockout MCF7 cell line (ab289293) at 20 µg
Lane 3:
Wild-type HeLa at 20 µg
Lane 4:
ARNTL knockout HeLa <a href='/en-us/products/cell-lines/human-arntl-bmal1-knockout-hela-cell-line-ab264701'>ab264701</a> at 20 µg
Lane 5:
Raji at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 69-75 kDa
Observed band size: 69-75 kDa
false
- NGS
Lab
Next Generation Sequencing - Human ARNTL knockout MCF7 cell line (AB289293)
103 bp deletion (allele 1) and 104 bp deletion (allele 2) after Thr294 of the WT protein.
Complementary Products
Primary Antibodies
AB230822
Anti-BMAL1 antibody [EPR23696-22]
BOND RX™ Validated|RabMAb|Recombinant|Advanced Validation|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-BMAL1 antibody [EPR23696-22] (AB230822)](https://content.abcam.com/products/images/bmal1-antibody-epr23696-22-ab230822--immunocytochemistry-immunofluorescence-img424968.jpg)
- 0
- 0
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cryptochrome I/CRY1 antibody [4H4-1C4] (AB54649)](https://content.abcam.com/products/images/cryptochrome-i-cry1-antibody-4h4-1c4-ab54649--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img94356.jpg)
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
Handling procedures
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.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
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.
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.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
MEM + 10% FBS + 0.01 mg/ml bovine insulin
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
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