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AB266975

Human REC8 knockout A549 cell line

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REC8 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 95 bp deletion in exon 6.

View Alternative Names

Cohesin Rec8p, Human homolog of rad21 S. pombe, Meiotic recombination and sister chromatid cohesion phosphoprotein of the rad21p family, Meiotic recombination protein REC8 homolog, Meiotic recombination protein REC8-like 1, REC8 homolog (yeast), REC8 like 1, REC8_HUMAN, Rec8p, Recombination and sister chromatid cohesion protein homolog

2 Images
Sanger Sequencing - Human REC8 knockout A549 cell line (AB266975)
  • Sanger seq

Unknown

Sanger Sequencing - Human REC8 knockout A549 cell line (AB266975)

Homozygous : 95 bp deletion in exon6

Cell Culture - Human REC8 knockout A549 cell line (AB266975)
  • Cell Culture

Unknown

Cell Culture - Human REC8 knockout A549 cell line (AB266975)

Representative images of REC8 knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 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 XL Core microscope.

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 95 bp deletion in exon 6

Antibiotic resistance

Puromycin 1µg/mL

Disease

Carcinoma

Product details

Recommended control: Human wild-type A549 cell line (ab255450). 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?

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Properties and storage information

Gene name
REC8
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 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.

The protein REC8 also known as Meiotic Recombination Protein REC8 Homolog plays a mechanical role in the cohesion of sister chromatids during meiosis. This protein weighs approximately 60 kDa and is expressed mostly in germ cells during the prophase of meiosis I. REC8 is structurally related to the cohesin complex family and its primary mechanical function is to maintain the physical connection between sister chromatids ensuring accurate chromosomal segregation during meiosis.
Biological function summary

Sister chromatid cohesion during meiosis depends heavily on REC8 which integrates into the cohesin protein complex. This complex consists of SMC1β SMC3 and STAG3 playing an essential role in meiotic chromosome pairing and segregation. REC8 forms part of this cohesin structure specifically ensuring precise cleavage and separation of chromatids during the anaphase I stage of meiosis which contributes critical fidelity in chromosomal inheritance.

Pathways

REC8 influences the meiotic cell cycle and the associated recombination pathways. These pathways are highly regulated and critical for gametogenesis. Within the meiotic cell cycle pathway REC8 interacts closely with the protein RAD21 which holds key roles across various stages of the cell cycle. The well-orchestrated removal of cohesion by separase enzyme ensures successful homologous chromosome separation highlighting REC8's importance in these cellular processes.

Defects in REC8 function show associations with infertility. Malfunctions in meiotic processes can lead to aneuploidies such as Down syndrome due to improper segregation of chromosomes. At the molecular level mutations or deficiencies in REC8 also connect to cohesinopathies like Cornelia de Lange syndrome where cohesion factor SMC3 could also play a part suggesting shared pathways or compensatory mechanisms related to chromosomal cohesion and segregation during meiosis.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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