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AB274948

Human GLO1 knockout MCF7 cell line

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GLO1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp deletion after Lys42 of the WT protein Frameshift = 100%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

Aldoketomutase, GLOD1, GLYI, Glx I, Glyoxalase I, Glyoxalase domain containing 1, Ketone-aldehyde mutase, LGUL_HUMAN, Lactoyl glutathione lyase, Methylglyoxalase, S-D-lactoylglutathione methylglyoxal lyase

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Next Generation Sequencing - Human GLO1 knockout MCF7 cell line (AB274948)
  • NGS

Lab

Next Generation Sequencing - Human GLO1 knockout MCF7 cell line (AB274948)

1 bp deletion after Lys42 of the WT protein

Key facts

Cell type

MCF7

Species or organism

Human

Tissue

Breast

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Mutation description

Knockout achieved by CRISPR/Cas9; X = 1 bp deletion after Lys42 of the WT protein Frameshift = 100%

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type MCF7 cell line (ab271144). 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.

Properties and storage information

Gene name
GLO1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage duration
1-2 weeks
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
  • Slow to trypsinise.
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 5-7x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

MEM + 10% FBS + 0.01 mg/ml bovine insulin

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 GLO1 protein also known as glyoxalase I plays an important role in cellular metabolism by detoxifying methylglyoxal a byproduct of glycolysis. It catalyzes the conversion of methylglyoxal into S-D-lactoylglutathione using glutathione as a cofactor. This enzyme has a mass of approximately 29 kDa and is expressed in various tissues throughout the body with higher levels found in the liver and kidney. Glyoxalase I (GLO1) expression is increased in response to stress conditions indicating its role in cellular defense mechanisms.
Biological function summary

Glyoxalase I is important for maintaining cellular homeostasis by mitigating harmful compounds. It forms part of the glyoxalase system which includes glyoxalase II functioning downstream of GLO1. This system prevents the accumulation of advanced glycation end-products (AGEs) which are implicated in cellular aging and dysfunction. By reducing the levels of methylglyoxal GLO1 supports normal physiological processes and protects cellular integrity.

Pathways

Glyoxalase I is an important component of the glyoxalase pathway. This pathway integrates into the detoxification network working alongside other important enzymes such as superoxide dismutase and catalase. These relationships highlight its function in cellular oxidative stress responses. GLO1's activity impacts pathways related to glycolysis and overall energy metabolism due to its role in reducing metabolic byproducts.

GLO1 has significant implications in diabetic complications and cancer. In diabetes increased methylglyoxal can worsen tissue damage making GLO1's role protective against advanced glycation end-product formation. In cancer GLO1 is overexpressed contributing to cellular proliferation and survival. Its interaction with proteins such as Hsp70 in cancer illustrates the adaptive mechanisms that tumor cells use for progression and chemoresistance.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

Product protocols

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