Human PDCD1 (PD1) knockout HeLa cell line
- Advanced Validation
- What is this?
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PDCD1 KO cell line available to order. KO validated. Free of charge wild type control available. 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. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 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] (<a href='/en-us/products/primary-antibodies/pd1-antibody-cal20-ab237728'>ab237728</a>) 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
false
- Sanger seq
Unknown
Sanger Sequencing - Human PDCD1 (PD1) knockout HeLa cell line (AB255417)
Allele-2 : 5 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human PDCD1 (PD1) knockout HeLa cell line (AB255417)
Allele-1 : 26 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human PDCD1 (PD1) knockout HeLa cell line (AB255417)
Allele-3 : Insertion of the selection cassette in exon 2.
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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Primary Antibodies
AB205921
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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