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AB287678

Human PDCD1 knockout A549 cell line

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PDCD1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

View Alternative Names

CD279 antigen, hPD 1, hPD l, hSLE1, PD 1, PD-1, PD1, PDCD 1, Programmed cell death 1, Programmed cell death 1 protein, Protein PD 1, SLEB2, Systemic lupus erythematosus susceptibility 2

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Next Generation Sequencing - Human PDCD1 knockout A549 cell line (AB287678)
  • NGS

Lab

Next Generation Sequencing - Human PDCD1 knockout A549 cell line (AB287678)

112 bp deletion after leu64

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.

What's included?

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Properties and storage information

Gene name
PDCD1
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.

PD1 also known as Programmed Cell Death Protein 1 or PDCD1 is a transmembrane protein that plays a critical role in regulating immune responses. It has a mass of approximately 55 kDa. PD1 is expressed on the surface of T cells B cells and some myeloid cells. PD1’s expression increases upon activation of these immune cells assisting in maintaining peripheral tolerance. Researchers often use PD1 mouse models and chimeric antibodies to explore the function of PD1 for experimental purposes. Antibodies such as anti-PD1 such as EH12.2H7 help in blocking PD1 interaction to study its role further.
Biological function summary

PD1 serves as an inhibitory receptor acting as a checkpoint in the immune system. It becomes part of an immune-suppressive complex when it binds with its ligands PD-L1 or PD-L2 which are expressed on various cell types including some tumor cells. This interaction suppresses the proliferation of T cells and cytokine production contributing to immune homeostasis. By controlling T cell activity PD1 limits autoimmunity but can also reduce the immune system's capability to attack cancer cells.

Pathways

PD1 functions in the immune checkpoint pathway a critical regulatory circuit in immune regulation. The engagement of PD1 with its ligands initiates a cascade that inhibits the function and proliferation of T cells through downstream SHP-2 phosphatase activity. This pathway frequently involves other regulatory proteins like CTLA-4 and is an important mechanism by which the body modulates immune responses. Related pathways often intersect with those involving T cell receptor signaling and contribute to the overall modulation of immune activity.

PD1 has a significant role in cancer and autoimmune disorders. PD1 expression can allow tumors to evade immune surveillance making PD1 a target for cancer therapies such as anti-PD1 antibodies which aim to block PD1 and restore T cell activity. The interaction of PD1 with cancer-related proteins like PD-L1 facilitates tumor immune evasion. In autoimmune disorders PD1’s regulation of immune balance can become dysregulated leading to persistent immune activation and tissue damage. Understanding PD1 and its interaction with proteins such as PD-L1 helps in developing therapeutic strategies for both cancer and autoimmune conditions.

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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