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AB258271

Human ULK3 knockout HEK-293T cell lysate

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ULK3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 1 bp insertion in exon1.

View Alternative Names

DKFZP434C131, FLJ90566, Serine/threonine-protein kinase ULK3, ULK3_HUMAN, Unc-51-like kinase 3, unc 51 like kinase 3 (C. elegans)

3 Images
Western blot - Human ULK3 knockout HEK-293T cell lysate (AB258271)
  • WB

Lab

Western blot - Human ULK3 knockout HEK-293T cell lysate (AB258271)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : ULK3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : HAP1 whole cell lyate (20 ug)
Lane 4 : LNCaP (Human prostate cancer cell line) whole cell lysate (20 ug)

ab124947 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 4oC 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 (<a href='/en-us/products/cell-lines/human-ulk3-knockout-hek-293t-cell-line-ab266152'>ab266152</a>)

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

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Sanger Sequencing - Human ULK3 knockout HEK-293T cell lysate (AB258271)
  • Sanger seq

Unknown

Sanger Sequencing - Human ULK3 knockout HEK-293T cell lysate (AB258271)

Allele-2 : 1 bp insertion in exon1

Sanger Sequencing - Human ULK3 knockout HEK-293T cell lysate (AB258271)
  • Sanger seq

Unknown

Sanger Sequencing - Human ULK3 knockout HEK-293T cell lysate (AB258271)

Allele-1 : 1 bp deletion in exon1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 1 bp insertion in exon1.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
ULK3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

ULK3 also known as Unc-51 Like Autophagy Activating Kinase 3 is a serine/threonine-protein kinase with a molecular mass of approximately 58 kDa. This protein plays an important role in mediating cellular processes through phosphorylation. ULK3 shows expression in various tissues including the brain and kidney. Its catalytic activity influences multiple cellular events especially those related to autophagy the cell's degradation pathway for removing damaged organelles and recycling cellular components.
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.

Researchers have reported associations with cancer and neurodegenerative diseases. Abnormal ULK3 activity can influence tumor growth and progression particularly in its regulatory role in cell proliferation and survival. Studies also suggest an involvement in neurodegenerative diseases where dysregulation in autophagy processes in which ULK3 plays a part contributes to neuronal cell death. In these contexts ULK3's interactions with proteins like mTOR and Beclin-1 may exacerbate disease progression by altering normal cellular processes.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB124947

Anti-ULK3 antibody [EPR4888]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ULK3 antibody [EPR4888] (AB124947)

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  • 0
Proteins & Peptides

AB101548

Recombinant human ULK3 protein

Functional Studies - Recombinant human ULK3 protein (AB101548)

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  • 0
Cell Lines & Lysates

AB266152

Human ULK3 knockout HEK-293T cell line

Advanced Validation

Western blot - Human ULK3 knockout HEK-293T cell line (AB266152)

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  • 0

Product promise

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For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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