Human ARNTL (BMAL1) knockout HeLa cell line
- Advanced Validation
- What is this?
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BMAL1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 10.
View Alternative Names
ARNT like protein 1 brain and muscle, Arntl, Aryl hydrocarbon receptor nuclear translocator like, Aryl hydrocarbon receptor nuclear translocator-like protein 1, BMAL1_HUMAN, BMAL1c, Basic helix loop helix PAS orphan MOP3, Basic-helix-loop-helix-PAS protein MOP3, Brain and muscle ARNT-like 1, CG8727 PA, Class E basic helix-loop-helix protein 5, JAP 3, MGC47515, MOP 3, Member of PAS protein 3, Member of PAS superfamily 3, PAS domain-containing protein 3, PASD 3, TIC, bHLH-PAS protein JAP3, bHLHe5, cycle
- 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.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
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
- 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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB230822
Anti-BMAL1 antibody [EPR23696-22]
BOND RX™ Validated|RabMAb|Recombinant|Advanced Validation|KO Validated
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- 0
- 0
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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