Human PRKD2 knockout A549 cell line
- Advanced Validation
- What is this?
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PRKD2 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 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
DKFZp586E0820, HSPC187, KPCD2_HUMAN, PRKD 2, Protein kinase D2, Serine/threonine-protein kinase D2, nPKC-D2
- WB
Lab
Western blot - Human PRKD2 knockout A549 cell line (AB301181)
Western blot : Anti-PRKD2 antibody (ab245527) staining at 1/3000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab245527 was shown to bind specifically to PRKD2. A band was observed at 105 kDa in wild-type A549 cell lysates with no signal observed at this size in PRKD2 knockout cell line. To generate this image, wild-type and PRKD2 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-PKD2 antibody (<a href='/en-us/products/primary-antibodies/pkd2-antibody-ab245527'>ab245527</a>) at 1/3000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
PRKD2 knockout A549 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 105 kDa
false
- WB
Lab
Western blot - Human PRKD2 knockout A549 cell line (AB301181)
Western blot : Anti-PRKD2 antibody [EP1495Y] (ab51250) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab51250 was shown to bind specifically to PRKD2. A band was observed at 105 kDa in wild-type A549 cell lysates with no signal observed at this size in PRKD2 knockout cell line. To generate this image, wild-type and PRKD2 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-PKD2 antibody [EP1495Y] (<a href='/en-us/products/primary-antibodies/pkd2-antibody-ep1495y-ab51250'>ab51250</a>) at 1/500 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
PRKD2 knockout A549 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 105 kDa
false
- NGS
Lab
Next Generation Sequencing - Human PRKD2 knockout A549 cell line (AB301181)
169 bp deletion after Asn 321 (allele 1); 180 bp deletion after Leu 319 (allele 2)
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
PKD2 plays a significant role in maintaining calcium ion homeostasis and cellular signal transduction. PKD2 functions together with PKD1 forming a complex that is essential in mechanosensation and ciliary signaling. When part of this complex PKD2 helps regulate cellular responses to mechanical stimuli which are important in tissue architecture and integrity. Furthermore PKD2 influences various cellular processes by modulating ion fluxes and intracellular signaling pathways.
Pathways
PKD2 is involved in the polycystic kidney disease pathway and the Wnt signaling pathway. These pathways are integral to cell proliferation differentiation and tissue maintenance. In the polycystic kidney disease pathway PKD2 interacts with PKD1 affecting cellular processes related to cyst formation. Additionally PKD2's role in the Wnt pathway which is vital for cellular communication and development highlights its cooperation with other signaling proteins like beta-catenin linking it to cellular growth and morphology.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Immunocytochemistry/ Immunofluorescence - Anti-PKD2 antibody [EP1495Y] (AB51250)](https://content.abcam.com/products/images/pkd2-antibody-ep1495y-ab51250--immunocytochemistry-immunofluorescence-img32960.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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