Human ATG16L1 knockout HeLa cell lysate (ab256843)
Overview
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Product name
Human ATG16L1 knockout HeLa cell lysate
See all ATG16L1 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 22 bp deletion in exon1 and Insertion of the selection cassette in exon1. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260157 - Human ATG16L1 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
Target
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Function
Plays an essential role in autophagy: interacts with ATG12-ATG5 to mediate the conjugation of phosphatidylethanolamine (PE) to LC3 (MAP1LC3A, MAP1LC3B or MAP1LC3C), to produce a membrane-bound activated form of LC3 named LC3-II. Thereby, controls the elongation of the nascent autophagosomal membrane. -
Involvement in disease
Inflammatory bowel disease 10 -
Sequence similarities
Belongs to the WD repeat ATG16 family.
Contains 7 WD repeats. -
Post-translational
modificationsProteolytic cleavage by activated CASP3 leads to degradation and may regulate autophagy upon cellular stress and apoptotic stimuli. -
Cellular localization
Cytoplasm. Preautophagosomal structure membrane. Recruited to omegasomes membranes by WIPI2. Omegasomes are endoplasmic reticulum connected strutures at the origin of preautophagosomal structures. Localized to preautophagosomal structure (PAS) where it is involved in the membrane targeting of ATG5. Localizes also to discrete punctae along the ciliary axoneme. - Information by UniProt
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Form
There are 4 isoforms produced by alternative splicing. -
Alternative names
- A16L1_HUMAN
- APG16 like 1
- APG16-like 1
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab256843 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa.
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| Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 68 kDa. |
Images
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Western blot - Human ATG16L1 knockout HeLa cell lysate (ab256843)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: ATG16L1 knockout HeLa cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: Daudi cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab233796 observed at 68 and 72 kDa. Red - loading control ab181602 observed at 37 kDa.
ab233796 Anti-ATG16L1 antibody [5H9A11] was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261772 (knockout cell lysate ab256843) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab233796 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Human ATG16L1 knockout HeLa cell lysate (ab256843)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: ATG16L1 knockout HeLa cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: Daudi cell lysate (20 µg)Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 68 and 72 kDa. Red - loading control ab8245 observed at 37 kDa.
ab187671 Anti-ATG16L1 antibody [EPR15638] - N-terminal was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261772 (knockout cell lysate ab256843) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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.
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Sanger Sequencing - Human ATG16L1 knockout HeLa cell lysate (ab256843)Allele-1: Insertion of the selection cassette in exon1
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Sanger Sequencing - Human ATG16L1 knockout HeLa cell lysate (ab256843)Allele-2: 22 bp deletion in exon1
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab256843 has not yet been referenced specifically in any publications.