Human PDE4B knockout HeLa cell line
- Advanced Validation
- What is this?
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PDE4B KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 12 and 1 bp insertion in exon 12.
View Alternative Names
5''-cyclic phosphodiesterase 4B, DKFZp686F2182, DPDE4, MGC126529, OTTHUMP00000010652, OTTHUMP00000010653, OTTHUMP00000010654, OTTHUMP00000010656, OTTHUMP00000232365, PDE IVB, PDE32, PDE4B5, PDE4B_HUMAN, Phosphodiesterase 4B, Phosphodiesterase 4B cAMP specific, Phosphodiesterase 4B, cAMP specific (phosphodiesterase E4 dunce homolog, Drosophila), cAMP-specific 3'', cAMP-specific 3',5'-cyclic phosphodiesterase 4B, cAMP-specific phosphodiesterase-4 B isoform, dunce-like phosphodiesterase E4
- WB
Lab
Western blot - Human PDE4B knockout HeLa cell line (AB265259)
Lanes 1-4 : Merged signal (red and green). Green - ab170939 observed at 83 kDa. Red - loading control ab8245 observed at 36 kDa.
ab170939 Anti-PDE4B antibody [EPR11830] was shown to specifically react with PDE4B in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265259 (knockout cell lysate ab257576) was used. Wild-type and PDE4B knockout samples were subjected to SDS-PAGE. ab170939 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-PDE4B antibody [EPR11830] (<a href='/en-us/products/primary-antibodies/pde4b-antibody-epr11830-ab170939'>ab170939</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PDE4B knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PDE4B knockout HeLa cell line (ab265259)
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
Mouse brain tissue lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 83 kDa
Observed band size: 83 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PDE4B knockout HeLa cell line (AB265259)
Allele-2 : 1 bp insertion in exon 12.
- Sanger seq
Unknown
Sanger Sequencing - Human PDE4B knockout HeLa cell line (AB265259)
Allele-1 : 19 bp deletion in exon 12.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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.
Culture medium
DMEM (High Glucose) + 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
PDE4B plays a significant role in modulating intracellular signaling by controlling the levels of cAMP a second messenger involved in various cellular processes. As part of the larger PDE4 family complex PDE4B assists in fine-tuning immune responses and inflammatory processes. This enzyme modulates the activity of protein kinase A (PKA) impacting processes such as cell proliferation differentiation and apoptosis.
Pathways
PDE4B operates within the cAMP signaling pathway which is essential for transducing extracellular signals into cellular responses. This pathway involves interaction with other proteins such as PKA and CREB (cAMP response element-binding protein) which are critical for regulating gene expression and cellular functions. PDE4B's ability to modulate this pathway makes it fundamental in controlling the dynamics of cellular signaling.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com