ANXA5 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
Alternative names
ANX 5, ANXA5_HUMAN, Anchorin CII, Annexin A5, Annexin V, Annexin-5, CBP-I, Calphobindin I, ENX-2, Endonexin II, Lipocortin V, PAP-I, PLACENTAL PROTEIN 4, PP 4, Placental anticoagulant protein 4, Placental anticoagulant protein I, RPRGL3, Thromboplastin inhibitor, VAC-alpha, Vascular anticoagulant-alpha
Recommended products
ANXA5 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
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- ANXA5
- 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
Annexin V also known as ANXA5 is a protein with a mass of approximately 35-36 kDa. This protein belongs to the annexin family which includes proteins capable of binding to phospholipids in a calcium-dependent manner. Annexin V is mostly expressed in placental tissue though it can be found in various other tissues like the vascular endothelium. Researchers often utilize annexin V for staining dead cells or identifying apoptosis using assays such as annexin V FITC or 7-AAD staining. It forms part of the annexin V binding buffer which enhances its binding affinity.
- Biological function summary
Annexin V has an important role in cell membrane dynamics and signaling. The protein does not function as part of a larger complex but its ability to bind negatively charged phospholipids like phosphatidylserine makes it important in apoptosis. During apoptosis phosphatidylserine translocates to the outer leaflet of the plasma membrane where annexin V marks these apoptotic cells. This property enables annexin V-based assays to detect early stages of apoptosis.
- Pathways
Annexin V involvement is seen in pathways associated with blood coagulation and apoptosis. In the apoptosis pathway annexin V interacts with apoptotic markers to identify dying cells. It relates to proteins such as caspases which are key executors of apoptosis. In coagulation annexin V has been shown to interact with prothrombinase complex and can affect calcium ion concentration thereby influencing blood clotting processes.
- Associated diseases and disorders
Annexin V plays a role in antiphospholipid syndrome and cardiovascular diseases. Annexin V has been researched for its connection to antiphospholipid antibodies where it affects normal annexin V function impacting clot formation. In cardiovascular diseases annexin V can interact with proteins such as thrombomodulin influencing thrombotic and atherosclerotic pathways. Understanding these interactions helps researchers develop potential therapeutic approaches for related conditions.
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3 product images
Western blot - Human ANXA5 knockout A549 cell line (ab300840)
Western blot: Anti-ANXA5 antibody [EPR3979] (Anti-Annexin V/ANXA5 antibody [EPR3979] ab108321) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Annexin V/ANXA5 antibody [EPR3979] ab108321 was shown to bind specifically to ANXA5. A band was observed at 36 kDa in wild-type A549 cell lysates with no signal observed at this size in ANXA5 knockout cell line. To generate this image, wild-type and ANXA5 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-Annexin V/ANXA5 antibody [EPR3979] (Anti-Annexin V/ANXA5 antibody [EPR3979] ab108321) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: Western blot - Human ANXA5 knockout A549 cell line (ab300840)
Lane 2: ANXA5 knockout A549 cell lysate at 20 µg
SecondaryLanes 1 - 2: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 36 kDa
Western blot - Human ANXA5 knockout A549 cell line (ab300840)
Western blot: Anti-ANXA5 antibody [EPR3980] (Anti-Annexin V/ANXA5 antibody [EPR3980] ab108194) staining at 1/10000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Annexin V/ANXA5 antibody [EPR3980] ab108194 was shown to bind specifically to ANXA5. A band was observed at 36 kDa in wild-type A549 cell lysates with no signal observed at this size in ANXA5 knockout cell line. To generate this image, wild-type and ANXA5 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-Annexin V/ANXA5 antibody [EPR3980] (Anti-Annexin V/ANXA5 antibody [EPR3980] ab108194) at 1/10000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: Western blot - Human ANXA5 knockout A549 cell line (ab300840)
Lane 2: ANXA5 knockout A549 cell lysate at 20 µg
SecondaryLanes 1 - 2: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 36 kDa
Next Generation Sequencing - Human ANXA5 knockout A549 cell line (ab300840)
47 bp deletion after Glu 71 of WT protein
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