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ARNTL 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.

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Images

Western blot - Human ARNTL (BMAL1) knockout HeLa cell line (AB264701), expandable thumbnail
  • Sanger Sequencing - Human ARNTL (BMAL1) knockout HeLa cell line (AB264701), expandable thumbnail

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

Knockout validation

Sanger Sequencing, Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 10

Alternative names

Recommended products

ARNTL 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.

Key facts

Cell type

HeLa

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 10

Disease

Adenocarcinoma

Concentration
Loading...

Properties

Gene name

ARNTL

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HeLa cell line (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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

2 product images

  • Western blot - Human ARNTL (BMAL1) knockout HeLa cell line (ab264701), expandable thumbnail

    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 Anti-GAPDH antibody [6C5] - 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 Human ARNTL (BMAL1) knockout HeLa 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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 3: SH-SY5Y cell lysate at 20 µg

    Lane 4: Huh7 cell lysate at 20 µg

    Performed under reducing conditions.

  • Sanger Sequencing - Human ARNTL (BMAL1) knockout HeLa cell line (ab264701), expandable thumbnail

    Sanger Sequencing - Human ARNTL (BMAL1) knockout HeLa cell line (ab264701)

    Homozygous: 1 bp deletion in exon 10.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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