PRKCD KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 2 bp deletion in exon 2.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 2 bp deletion in exon 2
CVID9, D14Ertd420e, KPCD_HUMAN, Kinase PKC delta, May-01, MGC49908, PCKd, PKC d, PKC delta, PRKC D, PRKC delta, Protein Kinase C delta, Protein kinase C delta VIII, Protein kinase C delta type, Tyrosine protein kinase PRKCD, nPKC-delta
PRKCD KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 2 bp deletion in exon 2.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 2 bp deletion in exon 2
G418 (Geneticin) 1µg/mL
PRKCD
Knockout
CRISPR technology
Sanger Sequencing
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.
This supplementary information is collated from multiple sources and compiled automatically.
Protein kinase C delta (PKC delta) often called PKCδ belongs to the novel PKC subfamily. It weighs approximately 78 kDa and functions in various cellular processes. This enzyme exhibits serine/threonine kinase activity and is expressed across numerous tissues including the brain heart and lymphoid organs. PKC delta activation mainly relies on diacylglycerol and phospholipids though it does not require calcium for activation. This target has multiple known phosphorylation sites such as delta 1485 and delta 647 that modulate its enzymatic activity and localization.
PKC delta influences cell growth apoptosis and differentiation through its kinase activity. It acts as an important regulator in the signaling pathways that control these processes. PKC delta is not a part of a traditional complex but interacts with several proteins influencing signaling cascades. In cellular contexts PKC delta positively influences tumor necrosis factor-related pathways and also initiates apoptosis in response to DNA damage.
PKC delta plays an essential role in the regulation of mitogen-activated protein kinase (MAPK) and Janus kinase (JAK) pathways. Within the MAPK pathway PKC delta interacts with proteins like c-Jun N-terminal kinase (JNK) facilitating stress-responsive pathways related to cell death and survival. In the JAK pathway its regulation can impact cytokine signaling and immune responses. PKC delta's function within these pathways positions it as an important mediator and influencer of diverse cellular responses.
PKC delta has significant implications for cancer and cardiovascular diseases. PKC delta promotes tumor cell proliferation in some cancers by altering cell cycle regulation and resist apoptosis. In cardiovascular disorders this protein mediates cardiac hypertrophy and heart failure through its influence on myocyte apoptosis and fibrosis. PKC delta’s interaction with specific proteins like Bcl-2 in cancer and troponin in cardiac conditions highlights its role in pathophysiological processes making it a significant target for therapeutic intervention.
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Lanes 1 - 2: Merged signal (red and green). Green - Anti-PKC delta antibody [EPR17075] ab182126 observed at 78 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-PKC delta antibody [EPR17075] ab182126 was shown to react with PKC in wild-type HEK-293T cells in western blot. The bands observed in PRKCD CRISPR/Cas9 edited cell line ab266143 (PRKCD CRISPR/Cas9 edited cell lysate Human PRKCD (PKC delta) knockout HEK-293T cell lysate ab257042) below 78kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and PRKCD CRISPR/Cas9 edited HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-PKC delta antibody [EPR17075] ab182126 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4&
176;C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-PKC delta antibody [EPR17075] (Anti-PKC delta antibody [EPR17075] ab182126) at 1/5000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: PRKCD CRISPR/Cas9 edited HEK-293T cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 78 kDa
Observed band size: 78 kDa
Allele-2: 1 bp insertion in exon 2.
Allele-1: 2 bp deletion in exon2
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