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AB257846

Human ATG4A knockout HeLa cell lysate

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ATG4A 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 exon4.
3 Images
Western blot - Human ATG4A knockout HeLa cell lysate (AB257846)
  • WB

Lab

Western blot - Human ATG4A knockout HeLa cell lysate (AB257846)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : ATG4A knockout HeLa cell lysate 20 μg
Lane 3 : HepG2 cell lysate 20 μg
Lane 4 : Daudi cell lysate 20 μg
Lanes 1 - 4 : Merged signal (red and green). Green - ab108322 observed at 45 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108322 was shown to react with ATG4A in wild-type HeLa cells in Western blot with loss of signal observed in ATG4A knockout cell line ab265738 (ATG4A knockout cell lysate ab257846). Wild-type HeLa and ATG4A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab108322 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-ATG4A antibody [EPR4122] (<a href='/en-us/products/primary-antibodies/atg4a-antibody-epr4122-ab108322'>ab108322</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ATG4A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ATG4A knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atg4a-knockout-hela-cell-line-ab265738'>ab265738</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 45 kDa

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Western blot - Human ATG4A knockout HeLa cell lysate (AB257846)
  • WB

Lab

Western blot - Human ATG4A knockout HeLa cell lysate (AB257846)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : ATG4A knockout HeLa cell lysate 20 μg
Lane 3 : HepG2 cell lysate 20 μg
Lane 4 : Daudi cell lysate 20 μg
Lanes 1 - 4 : Merged signal (red and green). Green - ab223374 observed at 45 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab223374 was shown to react with ATG4A in wild-type cells in Western blot with loss of signal observed in ATG4A knockout cell line ab265738 (ATG4A knockout cell lysate ab257846). Wild-type and ATG4A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab223374 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-ATG4A antibody (<a href='/en-us/products/primary-antibodies/atg4a-antibody-ab223374'>ab223374</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ATG4A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ATG4A knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atg4a-knockout-hela-cell-line-ab265738'>ab265738</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 45 kDa

Observed band size: 45 kDa

false

Sanger Sequencing - Human ATG4A knockout HeLa cell lysate (AB257846)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG4A knockout HeLa cell lysate (AB257846)

Homozygous : 1 bp deletion in exon4

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, 1 bp deletion in exon4.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB108322

Anti-ATG4A antibody [EPR4122]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG4A antibody [EPR4122] (AB108322)

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Cell Lines & Lysates

AB265738

Human ATG4A knockout HeLa cell line

Advanced Validation

Western blot - Human ATG4A knockout HeLa cell line (AB265738)

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Proteins & Peptides

AB172826

Recombinant Human ATG4A protein

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  • 0
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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 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
ATG4A
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.

ATG4A also known as Autophagin 1 is a cysteine protease with a mass of approximately 52 kDa. It plays a critical role in the autophagy pathway by cleaving LC3 and other Atg8 homologs facilitating their conjugation to phosphatidylethanolamine on the autophagosome membrane. This protein is expressed in various tissues including the liver and skeletal muscle indicating its involvement in diverse physiological processes.
Biological function summary

ATG4A contributes essential functions to cellular homeostasis by participating in autophagosome formation. As part of the group of Atg proteins it collaborates with other members to modulate the autophagy machinery. ATG4A's enzymatic activity provides the necessary steps for processing LC3 transforming it into a form that allows lipidation and incorporation into autophagic vesicles.

Pathways

ATG4A is integral to the autophagy pathway a catabolic process important for cell survival under stress conditions. It interacts with proteins such as ATG7 and ATG3 within this pathway playing a significant role in the early stages of autophagosome maturation. The interactions ensure proper recycling of cellular materials and its efficient function impact cellular metabolism and stress responses.

Disruptions in ATG4A function have been linked to neurodegenerative diseases and cancer. Aberrant autophagy processes involving ATG4A can lead to protein aggregation disorders such as Alzheimer’s disease where proper proteostasis is compromised. Also in certain cancers altered ATG4A activity can influence tumor growth often through modulation of metabolic pathways and interactions with proteins like Beclin 1 which regulate autophagic responses.
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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
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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