Human PML (PML Protein) knockout A549 cell line
- Advanced Validation
- What is this?
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PML KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 1 and 1 bp insertion in exon 1.
View Alternative Names
Acure promyelocytic leukemia, inducer of, MYL, PML_HUMAN, PP8675, Probable transcription factor PML, Promyelocytic leukemia, Promyelocytic leukemia inducer of, Promyelocytic leukemia protein, Protein PML, RING finger protein 71, RNF 71, TRIM 19, Tripartite motif protein TRIM19, Tripartite motif-containing protein 19
- WB
Lab
Western blot - Human PML (PML Protein) knockout A549 cell line (AB266980)
Lanes 1 - 2 : Merged signal (red and green). Green - ab179466 observed at 110 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab179466 was shown to react with PML in A549 wild-type cells in western blot with loss of signal observed in PML knockout cell line ab266980 (PML knockout cell lysate ab257082). Wild-type and PML knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk before incubation with ab179466 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-PML Protein antibody [EPR16792] (<a href='/en-us/products/primary-antibodies/pml-protein-antibody-epr16792-ab179466'>ab179466</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
PML knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human PML (PML Protein) knockout A549 cell line (ab266980)
Secondary
All lanes:
Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 110 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PML (PML Protein) knockout A549 cell line (AB266980)
Allele-2 : 1 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human PML (PML Protein) knockout A549 cell line (AB266980)
Allele-1 : 11 bp deletion in exon1
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
The PML protein interacts with various molecular partners and forms a part of multiprotein complexes within the PML-nuclear bodies. Its functions include the regulation of transcription induction of apoptosis DNA damage response and control of cell proliferation. PML can recruit other proteins such as p53 a tumor suppressor protein to influence these cellular activities. PML's ability to act as a scaffold within these complexes makes it essential for maintaining cellular homeostasis and response to stress. Various techniques such as PML protein ELISA are used to analyze PML-related biological activities.
Pathways
The PML protein plays a significant role in critical cellular pathways such as apoptosis and the interferon response pathway. PML's interaction with the p53 protein links it to the apoptosis pathway where it acts as an inducer of cell death in response to cellular stress and damage. In the interferon response pathway PML contributes to antiviral defense mechanisms. The involvement of PML in these pathways emphasizes its importance in cellular defense and programmed cell death. Related proteins like STATs (signal transducers and activators of transcription) are known to interact with PML in these pathways.
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
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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