PDCD1LG2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 22 bp insertion in exon 3.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 22 bp insertion in exon 3
Recommended products
PDCD1LG2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 22 bp insertion in exon 3.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 22 bp insertion in exon 3
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- PDCD1LG2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Cell culture
- Biosafety level
- EU: 1 US: 1
- Viability
- ~ 80%
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Cells should be passaged when they have achieved 80-90% confluence (approx 8x104-1x105 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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- 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 A549 cell line (ab255450). 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: F-12K + 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, 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
PD-L2 also known as PDCD1LG2 or B7-DC is a member of the B7 protein family that plays a role in immune system regulation. This protein has a molecular mass of approximately 25 kDa. PD-L2 is expressed in antigen-presenting cells such as dendritic cells and macrophages. Expression is also located in some subsets of T cells and B cells. The PD-L2 can act as a second ligand for the PD-1 receptor which is important in immune checkpoints.
- Biological function summary
PD-L2 offers protection to tissues from autoimmunity by regulating T-cell activation and fostering tolerance. By engaging with its receptor PD-1 PD-L2 forms a complex that modulates the immune response to prevent overactivation. This interaction limits the immune system's potential damage to healthy tissues maintaining proper immune function in diverse environments.
- Pathways
PD-L2 participates in the immune checkpoint pathways where it works closely with PD-1 to mediate immune responses. PD-L2 forms an important part of the program death-1 (PD-1) pathway which plays a prominent role in downregulating T-cell activity. It relates to proteins such as PD-L1 which also interacts with PD-1 modulating immune tolerance and response to infections or tumors.
- Associated diseases and disorders
PD-L2 involvement has been significant in cancer immunology and autoimmune conditions. Its interaction with PD-1 relates to the suppression of immune responses in tumors contributing to cancer progression. PD-L2's function as an immune checkpoint ligand associates with autoimmune diseases like rheumatoid arthritis where immune regulation goes awry. Both PD-1 and PD-L2 represent potential targets for therapeutic interventions due to their roles in these conditions.
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.
3 product images
Sanger Sequencing - Human PDCD1LG2 (PD-L2) knockout A549 cell line (ab267002)
Allele-1: 22 bp insertion in exon3
Sanger Sequencing - Human PDCD1LG2 (PD-L2) knockout A549 cell line (ab267002)
Allele-2: 1 bp deletion in exon 3.
Sanger Sequencing - Human PDCD1LG2 (PD-L2) knockout A549 cell line (ab267002)
Allele-3: 1 bp insertion in exon 3.
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