Human PRKCD (PKC delta) knockout HEK-293T cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
Western blot - Human PRKCD (PKC delta) knockout HEK-293T cell line (AB266143)
Lanes 1 - 2 : Merged signal (red and green). Green - ab182126 observed at 78 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
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 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 ab182126 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4<sup>&
176;</sup>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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/pkc-delta-antibody-epr17075-ab182126'>ab182126</a>) 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
Lane 2:
Western blot - Human PRKCD (PKC delta) knockout HEK-293T cell line (ab266143)
Predicted band size: 78 kDa
Observed band size: 78 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PRKCD (PKC delta) knockout HEK-293T cell line (AB266143)
Allele-1 : 2 bp deletion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human PRKCD (PKC delta) knockout HEK-293T cell line (AB266143)
Allele-2 : 1 bp insertion in exon 2.
Complementary Products
Primary Antibodies
AB182126
Anti-PKC delta antibody [EPR17075]
KO Validated|RabMAb|Recombinant|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-PKC delta antibody [EPR17075] (AB182126)](https://content.abcam.com/products/images/pkc-delta-antibody-epr17075-ab182126--immunocytochemistry-immunofluorescence-img123043.jpg)
- 4
- 7
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, 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
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.
Pathways
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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