NCF4 KO cell line available to order. Free of charge wild type control provided.
CGD3, MGC3810, NCF, NCF4_HUMAN, Neutrophil NADPH oxidase factor 4, Neutrophil cytosol factor 4, Neutrophil cytosolic factor 4, SH3 and PX domain-containing protein 4, SH3PXD4, p40-phox
NCF4 KO cell line available to order. Free of charge wild type control provided.
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.
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 HCT116 cell line (ab288559). 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.
P40-phox also known as neutrophil cytosolic factor 4 (NCF4) is a regulatory component of the NADPH oxidase complex. It is expressed in phagocytic cells such as neutrophils and macrophages. The protein plays a role in the production of reactive oxygen species (ROS) during the respiratory burst. The molecular mass of p40-phox is approximately 40 kDa. It interacts with other subunits of the NADPH oxidase complex such as p67-phox and p47-phox contributing to the assembly and activation of the complex.
P40-phox participates in innate immune responses by regulating the NADPH oxidase complex's function. It acts as a scaffold to facilitate the interaction between cytosolic and membrane components. p40-phox integrates into the cytosolic fraction with other subunits and translocates to the membrane upon stimulation. This translocation is essential for generating superoxide radicals that are involved in defense against pathogens. Its functional domains enable it to bind phosphatidylinositol (34) bisphosphate which anchors the protein to the membrane. p40-phox itself is not an enzyme but modulates the activity of the complex.
The p40 protein serves as an important player in the signaling cascade of the respiratory burst. It interacts with the phosphoinositide 3-kinase (PI3K) pathway and cooperates with other proteins like p67-phox and p47-phox through phosphorylation-dependent mechanisms. The p40-phox is phox compatible meaning it can integrate into various signaling modules. Within the oxidative burst pathway it enhances the cytosolic complex's attachment to the membrane affecting the production of ROS.
Mutations in the p40-phox have been connected to chronic granulomatous disease (CGD) characterized by recurrent infections and granuloma formation. The lack of functional p40-phox disrupts the assembly of the NADPH oxidase complex leading to decreased ROS production. This deficiency impairs the ability of phagocytes to eliminate pathogens effectively. p40-phox is also related to inflammatory disorders like inflammatory bowel disease (IBD) as variations in the NCF4 gene affect the immune response. The dysfunction in ROS production impacts the signaling involving p67-phox and p47-phox further influencing disease pathogenesis.
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