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AB257363

Human ATG3 knockout HEK-293T cell lysate

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ATG3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1.

View Alternative Names

2610016C12Rik, APG3 autophagy 3 like, APG3, S. cerevisiae, homolog of, APG3-like, APG3L, ATG3 autophagy related 3 homolog, ATG3 autophagy related 3 homolog (S. cerevisiae), ATG3_HUMAN, Apg 3, Apg3p, Autophagy 3, S. cerevisiae, homolog of, Autophagy Apg3p/Aut1p like, Autophagy-related protein 3, DKFZp564M1178, FLJ22125, MGC15201, OTTHUMP00000214547, OTTHUMP00000214548, PC3 96, Protein PC3-96, Ubiquitin-like-conjugating enzyme ATG3, autophagy related 3, hApg3

3 Images
Western blot - Human ATG3 knockout HEK-293T cell lysate (AB257363)
  • WB

Lab

Western blot - Human ATG3 knockout HEK-293T cell lysate (AB257363)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : ATG3 knockout HEK293T cell lysate (20 ug)
Lane 3 : K-562 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)

ab108251 was shown to specifically react with ATG3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266707 (knockout cell lysate ab257363) was used. Wild-type and ATG3 knockout samples were subjected to SDS-PAGE. ab108251 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-ATG3 antibody [EPR4801] (<a href='/en-us/products/primary-antibodies/atg3-antibody-epr4801-ab108251'>ab108251</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ATG3 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ATG3 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-atg3-knockout-hek-293t-cell-line-ab266707'>ab266707</a>)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

HeLa 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: 35 kDa

Observed band size: 40 kDa

false

Western blot - Human ATG3 knockout HEK-293T cell lysate (AB257363)
  • WB

Lab

Western blot - Human ATG3 knockout HEK-293T cell lysate (AB257363)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : ATG3 knockout HEK293T cell lysate (20 ug)
Lane 3 : K-562 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)

ab108282 was shown to specifically react with ATG3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266707 (knockout cell lysate ab257363) was used. Wild-type and ATG3 knockout samples were subjected to SDS-PAGE. ab108282 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-ATG3 antibody [EPR4802] (<a href='/en-us/products/primary-antibodies/atg3-antibody-epr4802-ab108282'>ab108282</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ATG3 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ATG3 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-atg3-knockout-hek-293t-cell-line-ab266707'>ab266707</a>)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

HeLa 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: 35 kDa

Observed band size: 40 kDa

false

Sanger Sequencing - Human ATG3 knockout HEK-293T cell lysate (AB257363)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG3 knockout HEK-293T cell lysate (AB257363)

Homozygous : 1 bp deletion in exon 1

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, Homozygous: 1 bp deletion in exon 1.

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Reactivity data

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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 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.

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What's included?

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Properties and storage information

Gene name
ATG3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

ATG3 also known as Autophagy-related protein 3 is an essential enzyme in the autophagy process. Its molecular weight is approximately 36 kDa. This protein facilitates the conjugation of LC3 (microtubule-associated protein 1A/1B-light chain 3) by acting as an E2-like enzyme. It is mainly expressed in tissues with high rates of autophagy including liver heart and brain.
Biological function summary

ATG3 plays an important role in the elongation of the autophagic membrane. It operates as a part of the autophagy machinery working closely with ATG7 an E1-like enzyme to transfer phosphatidylethanolamine (PE) to LC3 converting it into LC3-II. This lipidated form of LC3 is essential in the expansion and closure of the isolation membrane which forms the autophagosome the core structure for degradation and recycling of cellular components.

Pathways

ATG3 is a central player in the macroautophagy pathway a process critical for cellular homeostasis. It interacts with ATG7 to facilitate the lipidation of ATG8 family proteins which are important for autophagosome formation. Moreover ATG3 also links to apoptosis pathways where its balance with pro-apoptotic proteins helps regulate cell death processes.

ATG3 has been implicated in neurodegenerative diseases like Alzheimer's. This relates partly to its role in protein clearance which is essential for preventing toxic aggregations. Altered ATG3 function can lead to accumulation of misfolded proteins exacerbating disease. Moreover ATG3's interaction with proteins in the cancer-related pathways such as p53 shows its involvement in tumorigenesis through its influence on cell survival and death mechanisms.
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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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