Human CD14 knockout A549 cell line
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CD14 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
View Alternative Names
CD14 antigen, CD14 molecule, CD14_HUMAN, CD_antigen=CD14, LPS-R, Mo2, Monocyte differentiation antigen CD14, Monocyte differentiation antigen CD14 urinary form, Monocyte differentiation antigen CD14, membrane-bound form, Myeloid cell-specific leucine-rich glycoprotein
- NGS
Lab
Next Generation Sequencing - Human CD14 knockout A549 cell line (AB287269)
215 bp deletion after Asp78 (edit 1) and 103 bp deletion after Asp78 (edit 2)
Reactivity data
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?
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.
- 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.
Biological function summary
This glycoprotein plays a significant role in the innate immune system by recognizing pathogen-associated molecular patterns (PAMPs). It is part of a receptor complex where CD14 acts alongside LPS binding protein (LPS-R) TLR4 and MD-2 to facilitate pro-inflammatory signaling in response to microbial invasion. By binding to LPS CD14 initiates and amplifies the immune response leading to the production of cytokines and the recruitment of other immune cells to the site of infection.
Pathways
CD14 interacts with the toll-like receptor signaling pathway notably involving TLR4. Upon LPS detection CD14 assists in the activation of TLR4 triggering downstream signaling cascades such as NF-κB and MAPK pathways. These pathways result in the transcription of inflammatory cytokines and are essential for the host defense mechanism. The protein complex involving CD14 further interacts with adaptor proteins like MyD88 and TRIF facilitating signal propagation and the immune system's ability to respond to pathogens.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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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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