Human SQSTM1 (p62) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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SQSTM1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 4. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
A170, DMRV, EBI 3 associated protein p60, EBI3-associated protein of 60 kDa, EBIAP, FTDALS3, MGC127197, ORCA, OSF-6, OSIL, Osi, Oxidative stress induced like, P60, PDB 3, PKC-zeta-interacting protein, Paget disease of bone 3, Phosphotyrosine independent ligand for the Lck SH2 domain p62, Phosphotyrosine-independent ligand for the Lck SH2 domain of 62 kDa, Protein kinase C-zeta-interacting protein, SQSTM 1, SQSTM_HUMAN, STAP, STONE14, Sequestosome-1, Ubiquitin-binding protein p62, ZIP, ZIP 3, p62, p62B
- WB
Lab
Western blot - Human SQSTM1 (p62) knockout HEK-293T cell line (AB255343)
Lanes 1- 2 : Merged signal (red and green). Green - ab109012 observed at 64 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109012 was shown to react with SQSTM1 in wild-type HEK293Tcells in western blot. Loss of signal was observed when knockout cell line ab255343 (knockout cell lysate ab263770) was used. Wild-type HEK293Tand SQSTM1 knockout HEK293Tcell 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. ab109012 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 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-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker (<a href='/en-us/products/primary-antibodies/sqstm1-p62-antibody-epr4844-autophagosome-marker-ab109012'>ab109012</a>) at 1/10000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
SQSTM1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human SQSTM1 (p62) knockout HEK-293T cell line (ab255343)
Predicted band size: 47 kDa
Observed band size: 64 kDa
false
- WB
Lab
Western blot - Human SQSTM1 (p62) knockout HEK-293T cell line (AB255343)
Lanes 1- 2 : Merged signal (red and green). Green - ab56416 observed at 62 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab56416 was shown to react with SQSTM1 in wild-type HEK293Tcells in western blot. Loss of signal was observed when knockout cell line ab255343 (knockout cell lysate ab263770) was used. Wild-type HEK293Tand SQSTM1 knockout HEK293Tcell 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. ab56416 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 1000 Dilution and a 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.
All lanes:
Western blot - Anti-SQSTM1 / p62 antibody [2C11] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/sqstm1-p62-antibody-2c11-bsa-and-azide-free-ab56416'>ab56416</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
SQSTM1 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human SQSTM1 (p62) knockout HEK-293T cell line (ab255343)
Predicted band size: 47 kDa
Observed band size: 62 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SQSTM1 (p62) knockout HEK-293T cell line (AB255343)
Homozygous : 1 bp insertion in exon 4
- Sanger seq
Lab
Sanger Sequencing - Human SQSTM1 (p62) knockout HEK-293T cell line (AB255343)
Sequencing chromatogram displaying sequence edit in exon 4
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
Zygosity
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.
Quality control
STR analysis
D7S820, D5S818, TH01, D16S539, TPOX, CSF1PO, D13S317
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB109012
Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker
KO Validated|RabMAb|Recombinant|Lab Essentials
![Western blot - Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker (AB109012)](https://content.abcam.com/products/images/sqstm1-p62-antibody-epr4844-autophagosome-marker-ab109012--western-blot-img100339.jpg)
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Primary Antibodies
AB179483
Anti-HIF-1 alpha antibody [EPR16897]
RabMAb|Advanced Validation|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [EPR16897] (AB179483)](https://content.abcam.com/products/images/hif-1-alpha-antibody-epr16897-ab179483--immunocytochemistry-immunofluorescence-img85620.jpg)
- 4
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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