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AB257364

Human ATG4B knockout HeLa cell lysate

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ATG4B 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 and 1 bp insertion in exon4.
4 Images
Western blot - Human ATG4B knockout HeLa cell lysate (AB257364)
  • WB

Lab

Western blot - Human ATG4B knockout HeLa cell lysate (AB257364)

Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : ATG4B knockout HeLa cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab199537 observed at 47 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab199537 was shown to react with ATG4B in wild-type HeLa cells in Western blot with loss of signal observed in ATG4B knockout cell line ab260973 (ATG4B knockout cell lysate ab257364). Wild-type and ATG4B knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab199537 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-ATG4B antibody [EPR16572] (<a href='/en-us/products/primary-antibodies/atg4b-antibody-epr16572-ab199537'>ab199537</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ATG4B knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ATG4B knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atg4b-knockout-hela-cell-line-ab265814'>ab265814</a>)

Predicted band size: 44 kDa

Observed band size: 47 kDa

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Western blot - Human ATG4B knockout HeLa cell lysate (AB257364)
  • WB

Lab

Western blot - Human ATG4B knockout HeLa cell lysate (AB257364)

Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : ATG4B knockout HeLa cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab154843 observed at 47 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab154843 was shown to react with ATG4B in wild-type HeLa cells in Western blot with loss of signal observed in ATG4B knockout cell line ab260973 (ATG4B knockout cell lysate ab257364). Wild-type and ATG4B knockout HEK293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab154843 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-ATG4B antibody [EPR6436(2)] (<a href='/en-us/products/primary-antibodies/atg4b-antibody-epr64362-ab154843'>ab154843</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ATG4B knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ATG4B knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atg4b-knockout-hela-cell-line-ab265814'>ab265814</a>)

Predicted band size: 44 kDa

Observed band size: 47 kDa

false

Sanger Sequencing - Human ATG4B knockout HeLa cell lysate (AB257364)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG4B knockout HeLa cell lysate (AB257364)

Allele-1 : 1 bp deletion in exon4

Sanger Sequencing - Human ATG4B knockout HeLa cell lysate (AB257364)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG4B knockout HeLa cell lysate (AB257364)

Allele-2 : 1 bp insertion 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 and 1 bp insertion in exon4.

Disease

Adenocarcinoma

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Complementary Products

Proteins & Peptides

AB123195

Recombinant Human ATG4B protein

SDS-PAGE - Recombinant Human ATG4B protein (AB123195)

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  • 0
Primary Antibodies

AB154843

Anti-ATG4B antibody [EPR6436(2)]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-ATG4B antibody [EPR6436(2)] (AB154843)

  • 5
  • 2
Cell Lines & Lysates

AB265814

Human ATG4B knockout HeLa cell line

Advanced Validation

Western blot - Human ATG4B knockout HeLa cell line (AB265814)

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Cell Lines & Lysates

AB258762

Human WDR45 knockout HeLa cell lysate

Sanger Sequencing - Human WDR45 knockout HeLa cell lysate (AB258762)

  • 0
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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, 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
ATG4B
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.

ATG4B is a cysteine protease also known as autophagin-1 with a molecular mass of approximately 45 kDa. It is involved in the autophagy process by cleaving ATG8 family proteins. This protease facilitates the lipidation of LC3 an essential component of the autophagosome membrane. You find ATG4B expressed in various tissues especially in the brain liver and muscle where it plays important roles in cellular maintenance.
Biological function summary

ATG4B functions in the cellular autophagic machinery cleaving the C-terminal of ATG8 proteins. This processing is necessary for the conjugation to phosphatidylethanolamine and integration into autophagosomes. ATG4B is part of a larger autophagic complex that also includes other critical proteins like ATG7 and ATG3 helping prepare ATG8 proteins for autophagosome creation.

Pathways

ATG4B participates in the autophagy pathway by refining the cargo processing mechanism. It interacts with the mTOR signaling pathway which influences cell survival and growth based on nutrient availability. The ATG4B enzyme is closely related to ATG7 in the pathway as both contribute to the maturation of autophagosomes.

ATG4B has significant links to cancer and neurodegenerative disorders. In cancer ATG4B activity affects tumor survival since cancer cells exploit autophagy for growth under stress conditions. In neurodegenerative diseases like Alzheimer's alterations in ATG4B activity disrupt cellular homeostasis. This disruption connects to proteins like LC3 which are pivotal in neuron survival and function.
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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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