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CCND1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 1.

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Images

Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (AB255348), expandable thumbnail
  • Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (AB255348), expandable thumbnail
  • Sanger Sequencing - Human CCND1 (Cyclin D1) knockout HeLa cell line (AB255348), expandable thumbnail

Publications

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: 5 bp deletion in exon 1

Alternative names

Recommended products

CCND1 KO cell line available now. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 1.

Alternative names

Key facts

Cell type

HeLa

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 1

Disease

Adenocarcinoma

Concentration
Loading...

Properties

Gene name

CCND1

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 long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). 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.

3 product images

  • Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348), expandable thumbnail

    Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348)

    Lanes 1- 2: Merged signal (red and green). Green - ab40754 observed at 36 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

    ab40754 was shown to react with CCND1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate Human CCND1 (Cyclin D1) knockout HeLa cell lysate ab263808) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab40754 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    Lanes 1 - 2: Anti-Cyclin D1 antibody [EP272Y] - BSA and Azide free (ab227561) at 1/1000 dilution

    Lanes 1 - 2: Western blot - Anti-Cyclin D1 antibody [EP272Y] (ab40754) at 1/1000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: CCND1 knockout HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 34 kDa

    Observed band size: 36 kDa

  • Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348), expandable thumbnail

    Western blot - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348)

    Lanes 1- 2: Merged signal (red and green). Green - Anti-Cyclin D1 antibody [SP4] ab16663 observed at 36 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

    Anti-Cyclin D1 antibody [SP4] ab16663 was shown to react with CCND1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255348 (knockout cell lysate Human CCND1 (Cyclin D1) knockout HeLa cell lysate ab263808) was used. Wild-type HeLa and CCND1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Cyclin D1 antibody [SP4] ab16663 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 200 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Cyclin D1 antibody [SP4] (Anti-Cyclin D1 antibody [SP4] ab16663) at 1/200 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: CCND1 knockout HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 34 kDa

    Observed band size: 36 kDa

  • Sanger Sequencing - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348), expandable thumbnail

    Sanger Sequencing - Human CCND1 (Cyclin D1) knockout HeLa cell line (ab255348)

    Homozygous: 5 bp deletion in exon 1.

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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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