Human TP53 (p53) knockout A549 cell line
- Advanced Validation
- What is this?
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(3 Publications)
- WB
Lab
Western blot - Human TP53 (p53) knockout A549 cell line (AB276092)
False colour image of Western blot : Anti-p53 antibody [DO-1] - ChIP Grade staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab1101 was shown to bind specifically to p53. A band was observed at 40/50 kDa in treated wild-type A549 cell lysates with no signal observed at this size in tp53 knockout cell line ab276092. To generate this image, wild-type and tp53 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 (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
Lanes 1 - 12:
Western blot - Anti-p53 antibody [DO-1] (<a href='/en-us/products/primary-antibodies/p53-antibody-do-1-chip-grade-ab1101'>ab1101</a>) at 1/1000 dilution
Lanes 1 - 12:
Western blot - Anti-p53 antibody [DO-1] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/p53-antibody-do-1-bsa-and-azide-free-ab237976'>ab237976</a>)
Lane 1:
MCF7 Treated Camptothecin (20 ug/ml, 0 h) cell lysate at 15 µg
Lane 2:
MCF7 Treated Camptothecin (20 ug/ml, 6 h) cell lysate at 15 µg
Lane 3:
MCF7 Treated Camptothecin (20 ug/ml, 24 h) cell lysate at 15 µg
Lane 4:
Wild-type A549 Treated Camptothecin (20 ug/ml, 0 h) cell lysate at 15 µg
Lane 5:
Wild-type A549 Treated Camptothecin (20 ug/ml, 6 h) cell lysate at 15 µg
Lane 6:
Wild-type A549 Treated Camptothecin (20 ug/ml, 24 h) cell lysate at 15 µg
Lane 7:
TP53 knockout A549 Treated Camptothecin (20 ug/ml, 0 h) cell lysate at 15 µg
Lane 8:
TP53 knockout A549 Treated Camptothecin (20 ug/ml, 6 h) cell lysate at 15 µg
Lane 9:
TP53 knockout A549 Treated Camptothecin (20 ug/ml, 24 h) cell lysate at 15 µg
Lane 10:
Saos-2 cell lysate at 15 µg
Lane 11:
SK-BR-3 cell lysate at 15 µg
Lane 12:
A431 cell lysate at 15 µg
Secondary
All lanes:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution
false
- WB
Lab
Western blot - Human TP53 (p53) knockout A549 cell line (AB276092)
False colour image of Western blot : Anti-p53 antibody [DO-1] - ChIP Grade staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab1101 was shown to bind specifically to p53. A band was observed at 49 kDa in wild-type A549 cell lysates with no signal observed at this size in tp53 knockout cell line ab276092 (knockout cell lysate ab282999). To generate this image, wild-type and tp53 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 (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-p53 antibody [DO-1] (<a href='/en-us/products/primary-antibodies/p53-antibody-do-1-chip-grade-ab1101'>ab1101</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
Western blot - Human TP53 (p53) knockout A549 cell line (ab276092) at 20 µg
Lane 3:
Wild-type Jurkat cell lysate at 20 µg
Lane 4:
Western blot - Human TP53 (p53) knockout Jurkat cell line (<a href='/en-us/products/cell-lines/human-tp53-p53-knockout-jurkat-cell-line-ab276112'>ab276112</a>) at 20 µg
Lane 5:
A431 cell lysate at 20 µg
Lane 6:
Saos-2 cell lysate at 20 µg
Predicted band size: 43 kDa
Observed band size: 49 kDa
false
- WB
Lab
Western blot - Human TP53 (p53) knockout A549 cell line (AB276092)
False colour image of Western blot : Anti-p53 antibody [E26] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32389 was shown to bind specifically to p53. A band was observed at 49 kDa in wild-type A549 cell lysates with no signal observed at this size in tp53 knockout cell line ab276092 (knockout cell lysate ab282999). To generate this image, wild-type and tp53 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-p53 antibody [E26] (<a href='/en-us/products/primary-antibodies/p53-antibody-e26-ab32389'>ab32389</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
TP53 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human TP53 (p53) knockout Jurkat cell line (<a href='/en-us/products/cell-lines/human-tp53-p53-knockout-jurkat-cell-line-ab276112'>ab276112</a>)
Lane 3:
Wild-type Jurkat cell lysate at 20 µg
Lane 4:
TP53 knockout Jurkat cell lysate at 20 µg
Lane 5:
A431 cell lysate at 20 µg
Lane 6:
Saos-2 cell lysate at 20 µg
Predicted band size: 43 kDa
Observed band size: 49 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human TP53 (p53) knockout A549 cell line (AB276092)
49 bp deletion in exon 4
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Advanced Validation
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Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
Zygosity
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Publications (3)
Recent publications for all applications. Explore the full list and refine your search
Genome research 35:1456-1471 PubMed40360186
2025
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Species
Unspecified reactive species
Evidence-based complementary and alternative medicine : eCAM 2023:8378581 PubMed36814470
2023
Applications
Unspecified application
Species
Unspecified reactive species
Nature cell biology 24:1099-1113 PubMed35798843
2022
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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