Human PDCD6 (ALG-2) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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PDCD6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 1.
View Alternative Names
ALG-2-interacting protein 1, Apoptosis linked gene 2, Apoptosis-linked gene 2 protein, FLJ42309, FLJ46208, Hp95, KIAA1375, MA 3, MGC111017, MGC119050, MGC9123, PDCD6_HUMAN, PEF 1B, PS-2, Probable calcium-binding protein ALG-2, Programmed cell death 6, Programmed cell death 6-interacting protein, Programmed cell death protein 6
- WB
Lab
Western blot - Human PDCD6 (ALG-2) knockout HEK-293T cell line (AB266323)
Lanes 1-4 : Merged signal (red and green). Green - ab109181 observed at 22 kDa. Red - loading control ab8245 observed at 36 kDa.
ab109181 Anti-PDCD6/ALG-2 antibody [EP3259] was shown to specifically react with PDCD6/ALG-2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266323 (knockout cell lysate ab258570) was used. Wild-type and PDCD6/ALG-2 knockout samples were subjected to SDS-PAGE. ab109181 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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-PDCD6/ALG-2 antibody [EP3259] (<a href='/en-us/products/primary-antibodies/pdcd6-alg-2-antibody-ep3259-ab109181'>ab109181</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
PDCD6 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human PDCD6 (ALG-2) knockout HEK-293T cell line (ab266323)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Daudi cell 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: 21 kDa
Observed band size: 22 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PDCD6 (ALG-2) knockout HEK-293T cell line (AB266323)
Homozygous : 10 bp deletion in exon 1
- Cell Culture
Unknown
Cell Culture - Human PDCD6 (ALG-2) knockout HEK-293T cell line (AB266323)
Representative images of PDCD6 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Reactivity data
Product details
Recommended control: Human wild-type HEK293T 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 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
Zygosity
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
The function of PDCD6 involves its participation in calcium signaling pathways. This protein is part of a larger complex involving T-cell receptor signaling and also influences the Fas ligand-mediated apoptosis pathway. PDCD6 achieves its regulatory roles through interactions with various proteins such as MA3Ps. As a calcium-binding protein ALG-2 modulates calcium homeostasis which is critical for maintaining cellular functions including cell proliferation differentiation and programmed cell death.
Pathways
The involvement of PDCD6 in apoptosis is significant. It works in coordination with pathway proteins such as caspases and fas-associated proteins influencing cell death signaling. In addition PDCD6's ability to bind calcium allows it to fit into the broader intracellular signaling network placing it in connection with MAPK pathways essential for cellular responses to stress and growth signals. ALG protein interactions underline PDCD6's influence across apoptosis and signaling pathways.
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
Cell Lines & Lysates
AB266343
Human RAD23A (hHR23A) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
![Western blot - Anti-PDCD6/ALG-2 antibody [EPR3260] (AB133326)](https://content.abcam.com/products/images/pdcd6-alg-2-antibody-epr3260-ab133326--western-blot-img106801.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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