Human USP22 knockout HeLa cell lysate (ab257789)
Overview
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Product name
Human USP22 knockout HeLa cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1. -
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 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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Tested applications
Suitable for: Sanger Sequencing, WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260351 - Human USP22 knockout HeLa cell lysate 1 x 100µg ab255552 - 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
Histone deubiquitinating component of the transcription regulatory histone acetylation (HAT) complex SAGA. Catalyzes the deubiquitination of both histones H2A and H2B, thereby acting as a coactivator. Recruited to specific gene promoters by activators such as MYC, where it is required for transcription. Required for nuclear receptor-mediated transactivation and cell cycle progression. -
Tissue specificity
Moderately expressed in various tissues including heart and skeletal muscle, and weakly expressed in lung and liver. -
Sequence similarities
Belongs to the peptidase C19 family. UBP8 subfamily.
Contains 1 UBP-type zinc finger. -
Cellular localization
Nucleus. - Information by UniProt
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Alternative names
- Deubiquitinating enzyme 22
- KIAA1063
- Ubiquitin carboxyl terminal hydrolase 22
see all
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab257789 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 |
|---|---|---|
| Sanger Sequencing |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Predicted molecular weight: 60 kDa.
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| Notes |
|---|
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Sanger Sequencing
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 60 kDa. |
Images
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Western blot - Human USP22 knockout HeLa cell lysate (ab257789)
Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: USP22 knockout HeLa cell lysate 20 μg
False colour image of Western blot: Anti-USP22 antibody [EPR18945] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab195289 was shown to bind specifically to USP22. A band was observed at 59 kDa in wild-type HeLa cell lysates with no signal observed at this size in usp22 knockout cell line ab264888 (knockout cell lysate ab257789). To generate this image, wild-type and usp22 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. -
Western blot - Human USP22 knockout HeLa cell lysate (ab257789)Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: USP22 knockout HeLa cell lysate 20 μg
False colour image of Western blot: Anti-USP22 antibody [EPR4352(2)] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109435 was shown to bind specifically to USP22. A band was observed at 59 kDa in wild-type HeLa cell lysates with no signal observed at this size in usp22 knockout cell line ab264888 (knockout cell lysate ab257789). To generate this image, wild-type and usp22 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. -
Sanger Sequencing - Human USP22 knockout HeLa cell lysate (ab257789)Homozygous: 1 bp deletion in exon 1
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)
ab257789 has not yet been referenced specifically in any publications.