Human ULK3 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ULK3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1.
View Alternative Names
DKFZP434C131, FLJ90566, Serine/threonine-protein kinase ULK3, ULK3_HUMAN, Unc-51-like kinase 3, unc 51 like kinase 3 (C. elegans)
- WB
Lab
Western blot - Human ULK3 knockout HEK-293T cell line (AB266152)
Lanes 1-4 : Merged signal (red and green). Green - ab124947 observed at 55 kDa. Red - loading control ab8245 observed at 36 kDa.
ab124947 Anti-ULK3 antibody [EPR4888] was shown to specifically react with ULK3 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266152 (knockout cell lysate ab258271) was used. Wild-type and ULK3 knockout samples were subjected to SDS-PAGE. ab124947 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-ULK3 antibody [EPR4888] (<a href='/en-us/products/primary-antibodies/ulk3-antibody-epr4888-ab124947'>ab124947</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
ULK3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human ULK3 knockout HEK-293T cell line (ab266152)
Lane 3:
HAP1 whole cell lyate at 20 µg
Lane 4:
LNCaP (Human prostate cancer cell line) whole 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: 53 kDa
Observed band size: 55 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ULK3 knockout HEK-293T cell line (AB266152)
Allele-2 : 1 bp insertion in exon 1.
- Cell Culture
Lab
Cell Culture - Human ULK3 knockout HEK-293T cell line (AB266152)
Representative images ULK3 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 a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ULK3 knockout HEK-293T cell line (AB266152)
Allele-1 : 1 bp deletion in exon1
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, 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
This protein contributes significantly through its involvement in autophagy regulation. ULK3 acts as a kinase within a multiprotein complex which includes proteins such as ATG13 and FIP200. This complex is essential for initiating autophagy by regulating the autophagosome formation. Its activity can promote cellular survival under nutrient-deprived conditions showcasing its role as an important regulatory component in cellular maintenance and stress responses.
Pathways
ULK3 takes part in critical signaling pathways such as the mTOR signaling pathway and the Hedgehog signaling pathway. In the mTOR pathway ULK3 through its kinase activity acts upstream to regulate the activity of the autophagy machinery. It shares a functional relationship with proteins like mTOR and AMPK. In the Hedgehog pathway ULK3 regulates pathway components impacting cellular growth and differentiation processes. This positions ULK3 as a modulator connecting different signaling pathways to maintain cellular homeostasis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB124947
BOND RX™ Validated|RabMAb|Recombinant|KO Validated
![Western blot - Anti-ULK3 antibody [EPR4888] (AB124947)](https://content.abcam.com/products/images/ulk3-antibody-epr4888-ab124947--western-blot-img13111.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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