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AB289052

Human VTCN1 knockout A549 cell line

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VTCN1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

View Alternative Names

B7 family member, H4, B7 homolog 4, B7 superfamily member 1, B7S1, B7h.5, B7x, BC032925, Immune costimulatory protein B7-H4, MGC41287, PRO1291, Protein B7S1, RP11 229A19.4, T-cell costimulatory molecule B7x, V-set domain-containing T-cell activation inhibitor 1, VCTN1, VTCN1_HUMAN

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Next Generation Sequencing - Human VTCN1 knockout A549 cell line (AB289052)
  • NGS

Lab

Next Generation Sequencing - Human VTCN1 knockout A549 cell line (AB289052)

55 bp deletion after Pro 176 of WT protein

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Disease

Carcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

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 A549 cell line (ab288558). 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
VTCN1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation 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.
  • 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.

B7-H4 also known as VTCN1 or B7x is a member of the B7 family of immunoregulatory proteins. It has an approximate molecular mass of 50 kDa. B7-H4 expression is detected on the surface of various cells including B cells T cells monocytes and some cancer cells. Its expression is often inducible; immune cells can express B7-H4 in response to specific conditions or signals in the environment.
Biological function summary

B7-H4 acts as an immune checkpoint molecule that negatively regulates T cell immunity. It does not form a known complex with other proteins but instead interacts directly with T cells. By inhibiting T cell activation and proliferation B7-H4 contributes to the regulation of immune responses helping to maintain self-tolerance and prevent autoimmunity.

Pathways

B7-H4 is involved in the regulation of immune checkpoints prominently in the pathways related to immune tolerance and suppression. It plays a critical role in the pathways that modulate tumor evasion of the immune system. B7-H4 is related to other immune checkpoint proteins such as PD-L1 which are also pivotal in immune regulation and therapy targets.

B7-H4 is connected to cancer progression and autoimmune conditions. Overexpression of B7-H4 in certain cancers such as ovarian and breast cancer associates with tumor immune evasion allowing tumors to grow unchecked by the immune system. Its expression links with proteins like CTLA-4 in maintaining immune suppression. B7-H4 modifications could serve as a potential target in cancer immunotherapy aiming to enhance the immune system's ability to combat malignancies.

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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