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CD46 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.

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Images

Western blot - Human CD46 knockout A549 cell line (AB300878), expandable thumbnail
  • Next Generation Sequencing - Human CD46 knockout A549 cell line (AB300878), expandable thumbnail

Key facts

Cell type
A549
Species or organism
Human
Tissue
Lung
Form
Liquid
Knockout validation
Next Generation Sequencing, Western blot

Alternative names

AHUS2, Antigen defined by monoclonal antibody TRA 2 10, Antigen identified by monoclonal antibody TRA 2 10, CD46 antigen, CD46 antigen complement regulatory protein, CD46 molecule, CD46 molecule complement regulatory protein, Complement membrane cofactor protein, MCP_HUMAN, MGC26544, Measles virus receptor, Membrane cofactor protein, TLX, TRA2.10, Trophoblast leucocyte common antigen, Trophoblast leukocyte common antigen, Trophoblast lymphocyte cross reactive antigen, membrane cofactor protein (CD46, trophoblast-lymphocyte cross-reactive antigen)

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CD46 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.

Key facts

Cell type
A549
Form
Liquid
Disease
Carcinoma
Concentration
Loading...

Properties

Gene name
CD46
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot

Cell culture

Biosafety level
EU: 1 US: 1
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.
  • 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.
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 (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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

CD46 also known as membrane cofactor protein (MCP) is a type I membrane protein with a molecular weight around 50-70 kDa due to glycosylation variability. This protein serves as a regulatory component in the complement system which protects host cells from damage by complement components. CD46 is widely expressed on almost all nucleated human cells. It plays a role in inactivating C3b and C4b components of the complement cascade preventing autologous complement-mediated damage. The gene coding for CD46 is located on chromosome 1q32 a region known for encoding several complement regulatory proteins.

Biological function summary

Beyond its complement regulatory role CD46 acts in immune response modulation and reproduction. It forms a part of a protein complex that involves members such as decay-accelerating factor and complement receptor 1. CD46 also engages in intracellular signaling that affects T cell function and differentiation. In the reproductive context it participates in sperm-oocyte fusion and has a role in trophoblast fusion during placental formation.

Pathways

CD46 integrates into various signaling pathways including the complement and T cell receptor signaling pathways. In the complement pathway CD46 interacts with C3b and factor I promoting cleavage and inactivation of C3b leading to pathway regulation. Involvement in the T cell receptor pathway hints that CD46 signaling protects against overactive immune responses by promoting a switch to a regulatory T cell phenotype which can modulate immunity and prevent autoimmunity.

Associated diseases and disorders

CD46 has connections with autoimmune conditions and infectious diseases. For instance alterations in CD46 expression or function can contribute to atypical hemolytic uremic syndrome where dysregulation of the complement pathway occurs. Additionally certain pathogens like Neisseria gonorrhoeae utilize CD46 as a receptor for infection. CD46’s interaction with proteins such as C3b connects it directly to complement-related conditions highlighting its importance in maintaining immune system balance.

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.

2 product images

  • Western blot - Human CD46 knockout A549 cell line (ab300878), expandable thumbnail

    Western blot - Human CD46 knockout A549 cell line (ab300878)

    Western blot: Anti-CD46 antibody [EPR4014] (Anti-CD46 antibody [EPR4014] ab108307) staining at 1/1000 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 magenta. In Western blot, Anti-CD46 antibody [EPR4014] ab108307 was shown to bind specifically to CD46. A band was observed at 50-75 kDa in wild-type A549 cell lysates with no signal observed at this size in CD46 knockout cell line. To generate this image, wild-type and CD46 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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-CD46 antibody [EPR4014] (Anti-CD46 antibody [EPR4014] ab108307) at 1/1000 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: CD46 knockout A549 cell lysate at 20 µg

    Lane 3: HAP1 cell lysate at 20 µg

    Lane 4: THP-1 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

  • Next Generation Sequencing - Human CD46 knockout A549 cell line (ab300878), expandable thumbnail

    Next Generation Sequencing - Human CD46 knockout A549 cell line (ab300878)

    74 bp deletion after Pro 51 (allele 1); 1 bp insertion and 15 bp deletion after Tyr 52 (allele 2); 75 bp deletion after Pro 51 (allele 3); 1 bp insertion and 16 bp deletion after Tyr 52 (allele 4) of WT protein

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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