Human ANXA5 knockout A549 cell line
- Advanced Validation
- What is this?
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ANXA5 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- WB
Lab
Western blot - Human ANXA5 knockout A549 cell line (AB300840)
Western blot : Anti-ANXA5 antibody [EPR3980] (ab108194) staining at 1/10000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/annexin-v-anxa5-antibody-epr3980-ab108194'>ab108194</a>) 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
Secondary
Lanes 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
Observed band size: 36 kDa
false
- WB
Lab
Western blot - Human ANXA5 knockout A549 cell line (AB300840)
Western blot : Anti-ANXA5 antibody [EPR3979] (ab108321) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/annexin-v-anxa5-antibody-epr3979-ab108321'>ab108321</a>) 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
Secondary
Lanes 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
Observed band size: 36 kDa
false
- NGS
Lab
Next Generation Sequencing - Human ANXA5 knockout A549 cell line (AB300840)
47 bp deletion after Glu 71 of WT protein
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.
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
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.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Primary Antibodies
AB108194
Anti-Annexin V/ANXA5 antibody [EPR3980]
KO Validated|RabMAb|Recombinant
![Flow Cytometry (Intracellular) - Anti-Annexin V/ANXA5 antibody [EPR3980] (AB108194)](https://content.abcam.com/products/images/annexin-v-anxa5-antibody-epr3980-ab108194--flow-cytometry-intracellular-img179007.jpg)
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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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