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LAMTOR5 KO cell line available now. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.

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Images

Sanger Sequencing - Human LAMTOR5 (HBXIP) knockout HEK-293T cell line (AB266223), expandable thumbnail

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

Knockout validation

Sanger Sequencing

Mutation description

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

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LAMTOR5 KO cell line available now. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.

Key facts

Cell type

HEK-293T

Form

Liquid

Mutation description

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

Concentration
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Properties

Gene name

LAMTOR5

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing

Cell culture

Biosafety level

EU: 2 US: 2

Viability

~ 80%

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

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.

Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Culture medium: DMEM (High Glucose) + 10% FBS

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 2x10E4 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 2x10E4 cells/cm2 is recommended.

• Cells should be passaged when they have achieved 80-90% confluence.

• Do not allow the cell density to exceed 7x10E4 cells/cm2.

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.

Supplementary info

Activity summary

HBXIP also known as Hepatitis B X-interacting protein or LAMTOR5 is a protein with a mass of approximately 18 kDa. HBXIP binds to the HIV-1 protease and is involved in the activation of diverse cellular processes. It predominantly expresses in heart kidney liver and skeletal muscle tissues. The protein's mechanical role involves interacting with the hepatitis B virus X protein (HBx) enhancing the replication of hepatitis B virus particles.

Biological function summary

HBXIP impacts multiple cellular functions in a complex manner. It participates in the regulation of centrosome dynamics and cell division. HBXIP interacts with proteins such as survivin in a complex that plays a role in mitosis. HBXIP also influences gene expression by modulating estrogen receptor signaling. These functions make HBXIP a significant component in cellular growth control.

Pathways

HBXIP engages in cell cycle regulation and apoptosis. It affects the mTOR signaling pathway important for cell growth and metabolism. Within this context HBXIP acts alongside proteins like mTOR and RHEB influencing nutrient and energy status in the cell. Furthermore its interaction with the p53 pathway highlights its role in controlling cell survival and the response to DNA damage.

Associated diseases and disorders

HBXIP displays connections to cancer and viral infections. In cancer HBXIP overexpression correlates with tumor progression and poor patient prognosis commonly seen in breast cancer. Through its influence on nuclear factor-kB (NF-kB) HBXIP associates with inflammatory responses tied to cancer progression. Additionally its relationship with hepatitis B viral oncogenesis highlights a link between HBXIP and liver disorders signaling interaction with the viral HBx protein as a point of mechanism.

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.

1 product image

  • Sanger Sequencing - Human LAMTOR5 (HBXIP) knockout HEK-293T cell line (ab266223), expandable thumbnail

    Sanger Sequencing - Human LAMTOR5 (HBXIP) knockout HEK-293T cell line (ab266223)

    Homozygous: 1 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'.

For licensing inquiries, please contact partnerships@abcam.com