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AB258786

Human ATG10 knockout HeLa cell lysate

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ATG10 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 5 bp deletion in exon5 and 92 bp insertion in exon5.

View Alternative Names

APG 10L, APG10 autophagy 10 like, APG10, S. cerevisiae, homolog of, APG10-like, ATG10 autophagy related 10 homolog, ATG10 autophagy related 10 homolog (S. cerevisiae), ATG10_HUMAN, Apg 10, Autophagy-related protein 10, DKFZP586I0418, FLJ13954, Ubiquitin-like-conjugating enzyme ATG10, autophagy 10, S. cerevisiae, homolog of, pp12616

3 Images
Western blot - Human ATG10 knockout HeLa cell lysate (AB258786)
  • WB

Lab

Western blot - Human ATG10 knockout HeLa cell lysate (AB258786)

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 2 : ATG10 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 3 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate (20 ug)
Lane 4 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)

ab124711 was shown to specifically react with ATG10 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265682 (knockout cell lysate ab258786) was used. Wild-type and ATG10 knockout samples were subjected to SDS-PAGE. ab124711 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-ATG10 antibody [EPR4804] (<a href='/en-us/products/primary-antibodies/atg10-antibody-epr4804-ab124711'>ab124711</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

ATG10 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human ATG10 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atg10-knockout-hela-cell-line-ab265682'>ab265682</a>)

Lane 3:

K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) 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: 25 kDa

Observed band size: 28 kDa

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Sanger Sequencing - Human ATG10 knockout HeLa cell lysate (AB258786)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG10 knockout HeLa cell lysate (AB258786)

Allele-2 : 5 bp deletion in exon5

Sanger Sequencing - Human ATG10 knockout HeLa cell lysate (AB258786)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG10 knockout HeLa cell lysate (AB258786)

Allele-1 : 92 bp insertion in exon5

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 5 bp deletion in exon5 and 92 bp insertion in exon5.

Disease

Adenocarcinoma

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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
ATG10
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.

ATG10 also known as Autophagy-Related Protein 10 is an important player in the autophagy pathway. This protein weighs approximately 21 kDa and is expressed in a variety of tissues with high levels observed in the heart and skeletal muscle. Mechanically ATG10 acts as an E2-like enzyme important for the conjugation of ATG12 to ATG5 forming the ATG12–ATG5 conjugate essential for autophagosome formation.
Biological function summary

The autophagy process involves ATG10 in the covalent attachment of ATG12 to ATG5 a step critical for elongation and completion of autophagosomes. This protein functions as part of the ATG12–ATG5–ATG16L1 complex which plays a structural role in the phagophore membrane. Without ATG10's activity efficient autophagosome assembly cannot occur highlighting its importance in maintaining cellular homeostasis through regulated turnover of cellular components.

Pathways

ATG10 significantly impacts the macroautophagy pathway and links to metabolic control pathways. It interacts closely with proteins such as ATG5 and ATG7 which are necessary for initiating and executing the autophagy process. This involvement ensures the proper degradation of damaged organelles and recycling of nutrients during cellular stress.

Researchers have connected ATG10 activity to conditions like neurodegenerative diseases and cancers. Dysregulation of ATG10 can impair autophagic function which may contribute to Alzheimer's disease progression by preventing the clearance of protein aggregates. Additionally alteration in ATG10 expression has associations with certain cancers where it impacts cancer cell survival and proliferation. Understanding ATG10's interactions with other proteins like BECN1 could further elucidate its role in these diseases.
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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

AB247975

Anti-ATG10 antibody [EPR4804] - BSA and Azide free

RabMAb|Recombinant|KO Validated

Western blot - Anti-ATG10 antibody [EPR4804] - BSA and Azide free (AB247975)

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Cell Lines & Lysates

AB265682

Human ATG10 knockout HeLa cell line

Advanced Validation

Western blot - Human ATG10 knockout HeLa cell line (AB265682)

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Primary Antibodies

AB124711

Anti-ATG10 antibody [EPR4804]

RabMAb|Recombinant|KO Validated

Western blot - Anti-ATG10 antibody [EPR4804] (AB124711)

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Primary Antibodies

AB62352

Anti-Nrf2 antibody [EP1808Y]

RabMAb|Recombinant|KO Validated

Western blot - Anti-Nrf2 antibody [EP1808Y] (AB62352)

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