Human USP9X knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human USP9X knockout HeLa cell line (AB265665)
Lanes 1- 2 : Merged signal (red and green). Green - ab19879 observed at 290 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab19879 was shown to react with USP9x in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265665 (knockout cell lysate ab257790) was used. Wild-type HeLa and USP9X knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab19879 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 μg/ml and a 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-USP9x antibody (<a href='/en-us/products/primary-antibodies/usp9x-antibody-ab19879'>ab19879</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
USP9X knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human USP9X knockout HeLa cell line (ab265665)
Predicted band size: 292 kDa
Observed band size: 290 kDa
false
- WB
Lab
Western blot - Human USP9X knockout HeLa cell line (AB265665)
Lanes 1- 2 : Merged signal (red and green). Green - ab180191 observed at 290 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab180191 was shown to react with USP9x in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265665 (knockout cell lysate ab257790) was used. Wild-type HeLa and USP9X knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180191 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 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-USP9x antibody [EPR13809(B)] - N-terminal (<a href='/en-us/products/primary-antibodies/usp9x-antibody-epr13809b-n-terminal-ab180191'>ab180191</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
USP9X knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human USP9X knockout HeLa cell line (ab265665)
Predicted band size: 140 kDa,292 kDa
Observed band size: 220 kDa,290 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human USP9X knockout HeLa cell line (AB265665)
Allele-2 : 1 bp insertion in exon 9.
- Sanger seq
Unknown
Sanger Sequencing - Human USP9X knockout HeLa cell line (AB265665)
Allele-1 : 1 bp deletion in exon 9.
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Cell Lines & Lysates
AB265788
Human PPID (Cyclophilin 40) knockout HeLa cell line
Advanced Validation
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Primary Antibodies
AB180191
Anti-USP9x antibody [EPR13809(B)] - N-terminal
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-USP9x antibody [EPR13809(B)] - N-terminal (AB180191)](https://content.abcam.com/products/images/usp9x-antibody-epr13809b-n-terminal-ab180191--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img84391.jpg)
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Cell Lines & Lysates
AB264902
Human PHF6 (PHD finger protein 6) knockout HeLa cell line
Advanced Validation
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Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
USP9x influences numerous cellular mechanisms by maintaining protein balance. It is a part of the complex cellular machinery involved in signal transduction cell division and apoptosis. While its enzyme activity regulates various cellular proteins USP9x is essential in neural development and differentiation. The proteolytic activity helps to modulate pathways by controlling protein turnover which impacts cellular responses and development. Failure to regulate proteins accurately can lead to compromised cellular functions.
Pathways
USP9x participates prominently in the Wnt signaling and TGF-beta pathways. Its role in deubiquitination allows it to stabilize proteins like beta-catenin and SMAD4 which are vital for transmitting signals within these pathways. Through these interactions USP9x aids in cell proliferation migration and differentiation often connecting with other proteins like APC or Axin in the Wnt pathway. This demonstrates how USP9x's enzymatic activity intricately links to key biological signaling mechanisms.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com