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AB265749

Human RAD1 knockout HeLa cell line

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RAD1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 2 bp deletion in exon 2 and 62 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Sanger Sequencing - Human RAD1 knockout HeLa cell line (AB265749)
  • Sanger seq

Unknown

Sanger Sequencing - Human RAD1 knockout HeLa cell line (AB265749)

Allele-2 : 1 bp insertion in exon 2.

Sanger Sequencing - Human RAD1 knockout HeLa cell line (AB265749)
  • Sanger seq

Unknown

Sanger Sequencing - Human RAD1 knockout HeLa cell line (AB265749)

Allele-3 : 62 bp deletion in exon 2.

Sanger Sequencing - Human RAD1 knockout HeLa cell line (AB265749)
  • Sanger seq

Unknown

Sanger Sequencing - Human RAD1 knockout HeLa cell line (AB265749)

Allele-1 : 2 bp deletion in exon 2.

Cell Culture - Human RAD1 knockout HeLa cell line (AB265749)
  • Cell Culture

Lab

Cell Culture - Human RAD1 knockout HeLa cell line (AB265749)

Representative images RAD1 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 2 bp deletion in exon 2 and 62 bp deletion in exon 2

Disease

Adenocarcinoma

Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

What's included?

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Properties and storage information

Gene name
RAD1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Rad1 also known as RAD1 homolog 1 from Saccharomyces cerevisiae functions mechanically as an exonuclease involved in DNA repair processes. It forms a part of the 9-1-1 checkpoint complex which acts as a sensor for DNA damage. Rad1 has a molecular mass of approximately 34 kDa and is expressed in various tissues where cell division occurs. Its primary role is in maintaining genomic stability by participating in the recognition and repair of abnormal DNA structures.
Biological function summary

Rad1 plays an essential role in the DNA damage response by being part of the 9-1-1 complex which includes Rad9 and Hus1 proteins. This complex is an important player in activating the ATR signaling pathway which helps cells respond to DNA replication stress and damage. Rad1 helps ensure proper checkpoint activation preventing cells with damaged DNA from progressing through the cell cycle which further safeguards genomic integrity.

Pathways

The protein Rad1 functions critically within the ATR-Chk1 signaling pathway a significant route for controlling cell cycle checkpoints. In this context Rad1 associates with proteins such as ATR and Chk1 facilitating the signaling necessary for cell cycle arrest under conditions of DNA stress. Rad1 also interacts with the base excision repair pathway where it works with other repair proteins to rectify oxidative DNA damage and maintain cellular viability.

Rad1 has a connection with cancer due to its involvement in DNA repair pathways. Impaired Rad1 function can lead to accumulation of DNA damage contributing to carcinogenesis. Additionally Rad1's participation in maintaining genomic stability links it to disorders like Fanconi anemia where DNA repair defects are evident. In these contexts Rad1's interactions with proteins like BRCA1 further highlight its importance in protective cellular mechanisms.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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

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