Human wild-type A549 cell line is not available for individual purchase. This product can be added free of charge to a KO cell line order of the same background.
Images
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
Recommended products
Human wild-type A549 cell line is not available for individual purchase. This product can be added free of charge to a KO cell line order of the same background.
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Disease
Carcinoma
- Concentration
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.
- 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.
Storage
- Shipped at conditions
Dry Ice
- Appropriate short-term storage conditions
-196°C
- Appropriate long-term storage conditions
-196°C
Notes
Wild-type cell lines are sold with knockout cell lines only - not available for individual purchase.
We will provide viable cells that proliferate on revival.
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 wild-type A549 cell line (ab288558)
Western blot: Anti-TERT antibody staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab52866) loading control staining at 1/20000 dilution, shown in magenta.
In Western blot, the antibody was shown to bind specifically to TERT. A band was observed at 105 kDa in wild-type A549 cell lysates with no signal observed at this size in TERT knockout cell line Human TERT knockout A549 cell line ab289032.
To generate this image, wild-type and TERT 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Anti-TERT antibody at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate (ab288558) at 20 µg
Lane 2: TERT knockout A549 cell lysate (Human TERT knockout A549 cell line ab289032) at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: U-2 OS cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Western blot - Human wild-type A549 cell line (ab288558)
False colour image of Western blot: Anti-BRD2 antibody [BL-167-2A2] staining at 1/1000 dilution, shown in green; Mouse Anti-Nucleophosmin antibody [FC82291] (Anti-Nucleophosmin antibody [FC82291] ab10530) loading control staining at 2 ug/mL imaged in ECL. In Western blot, Anti-BRD2 antibody [BL-167-2A2] ab243865 was shown to bind specifically to BRD2. A band was observed at 110 kDa in wild-type A549 cell lysates with no signal observed at this size in BRD2 knockout cell line. To generate this image, wild-type and BRD2 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 before development with Optiblot (ECL reagent Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaged with 20 seconds exposure time. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and HRP conjugated Goat anti-Mouse (H+L) at 1/20000 dilution.
All lanes: Western blot - Anti-BRD2 antibody [BL-167-2A2] (Anti-BRD2 antibody [BL-167-2A2] ab243865) at 1/1000 dilution
Lane 1: Wild-type A549 cytoplasm cell lysate at 20 µg
Lane 2: Wild-type A549 nuclear cell lysate at 20 µg
Lane 3: BRD2 knockout A549 C7 cytoplasm cell lysate at 20 µg
Lane 4: BRD2 knockout A549 C7 nuclear cell lysate at 20 µg
Lane 5: Wild-type HEK-293T cell lysate at 20 µg
Lane 6: BRD2 knockout HEK-293T cell lysate at 20 µg
Performed under reducing conditions.
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