PDCD1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 26 bp deletion in exon 2 and 5 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 26 bp deletion in exon 2 and 5 bp deletion in exon 2 and Insertion of the selection cassette in exon 2
Alternative names
CD279, CD279 antigen, PD-1, PDCD1_HUMAN, Programmed cell death 1, Programmed cell death 1 protein, Programmed cell death protein 1, Protein PD-1, SLEB2, Systemic lupus erythematosus susceptibility 2, hPD-1, hSLE1
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PDCD1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 26 bp deletion in exon 2 and 5 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 26 bp deletion in exon 2 and 5 bp deletion in exon 2 and Insertion of the selection cassette in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- PDCD1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- 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 short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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4 product images
Western blot - Human PDCD1 (PD1) knockout HeLa cell line (ab255417)
False colour image of Western blot: Anti-PD1 antibody [CAL20] staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-PD1 antibody [CAL20] ab237728 was shown to bind specifically to PD1. A band was observed at 48 kDa in wild-type HeLa cell lysates with no signal observed at this size in PDCD1 CRISPR-Cas9 edited cell line ab255417 (CRISPR-Cas9 edited cell lysate Human PDCD1 (PD1) knockout HeLa cell lysate ab263794). The band observed in the CRISPR-Cas9 edited lysate lane below 48 kDa is likely to represent a truncated form of PD1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and PDCD1 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-PD1 antibody [CAL20] (Anti-PD1 antibody [CAL20] ab237728) at 1/500 dilution
Lane 1: Wild-type HeLa Vehicle Control lonomycin (0 ng/mL, 24h) + PMA (10 ng/mL, 24 h) cell lysate at 20 µg
Lane 2: Wild-type HeLa Treated lonomycin (500 ng/mL, 24h) + PMA (10 ng/mL, 24 h) cell lysate at 20 µg
Lanes 3 - 4: Western blot - Human PDCD1 (PD1) knockout HeLa cell line (ab255417)
Lane 3: PDCD1 knockout HeLa vehicle Control lonomycin (0ng/mL 24h) + PMA (10ng/mL 24h) cell lysate at 20 µg
Lane 4: PDCD1 knockout HeLa lonomycin (500ng/mL 24h) + PMA (10ng/mL 24h) cell lysate at 20 µg
Performed under reducing conditions.
Sanger Sequencing - Human PDCD1 (PD1) knockout HeLa cell line (ab255417)
Allele-2: 5 bp deletion in exon 2.
Sanger Sequencing - Human PDCD1 (PD1) knockout HeLa cell line (ab255417)
Allele-3: Insertion of the selection cassette in exon 2.
Sanger Sequencing - Human PDCD1 (PD1) knockout HeLa cell line (ab255417)
Allele-1: 26 bp deletion in exon 2.
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