Human ARNTL (BMAL1) knockout HeLa cell line
- Advanced Validation
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BMAL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 10. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Aryl hydrocarbon receptor nuclear translocator-like protein 1, bHLHe5, Member of PAS protein 3, MOP3, BMAL1, Basic-helix-loop-helix-PAS protein MOP3, ARNTL, PAS domain-containing protein 3, PASD3, BHLHE5, Brain and muscle ARNT-like 1, Basic helix-loop-helix ARNT-like protein 1, bHLH-PAS protein JAP3, Class E basic helix-loop-helix protein 5
- WB
Lab
Western blot - Human ARNTL (BMAL1) knockout HeLa cell line (AB264701)
Lanes 1 - 4 : Merged signal (red and green). Green - Anti-ß1-Adrenergic Receptor antibody observed at 76 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
Anti-ß1-Adrenergic Receptor antibody was shown to react with ß1-Adrenergic Receptor in wild-type HeLa cells in Western blot with loss of signal observed in ARNTL knockout cell line ab264701 (ARNTL knockout cell lysate ab258314). Wild-type HeLa and ARNTL knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-ß1-Adrenergic Receptor antibody and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Lane 1:
Anti-ß1-Adrenergic Receptor antibody at 1/1000 dilution
Lanes 2 - 4:
Anti-β1-Adrenergic Receptor antibody at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ARNTL knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ARNTL (BMAL1) knockout HeLa cell line (ab264701)
Lane 3:
SH-SY5Y cell lysate at 20 µg
Lane 4:
Huh7 cell lysate at 20 µg
false
- Sanger seq
Unknown
Sanger Sequencing - Human ARNTL (BMAL1) knockout HeLa cell line (AB264701)
Homozygous : 1 bp deletion in exon 10.
- WB
Lab
Western blot - Human ARNTL (BMAL1) knockout HeLa cell line (AB264701)
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 1/20000 dilution, shown in magenta. A band was observed at 69 kDa in Wild-type A549 and HeLa cell lysates with no signal observed at this size in ARNTL knockout A549 cell line and ARNTL knockout HeLa cell line. To generate this image, 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 800CW and 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 A549 cell lysates at 20 µg
Lane 2:
Western blot - Human ARNTL knockout A549 cell line (ab288835) at 20 µg
Lane 3:
Wild-type HeLa ab255928 cell lysates at 20 µg
Lane 4:
Western blot - Human ARNTL (BMAL1) knockout HeLa cell line (ab264701) at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 69 kDa
Observed band size: 69 kDa
false
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
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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