Human SQSTM1 (p62) knockout HCT116 cell line
- Advanced Validation
- What is this?
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SQSTM1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 49 bp deletion 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 HCT116 cell line (AB266871)
Lanes 1-4 : Merged signal (red and green). Green - ab109012 observed at 55 kDa. Red - loading control ab8245 observed at 36 kDa.
ab109012 Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker was shown to specifically react with SQSTM1 / p62 in wild-type HCT116 cells. The band observed in CRISPR/Cas9 edited cell line ab266871 (CRISPR/Cas9 edited cell lysate ab257052) lane below 55 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and SQSTM1 / p62 CRISPR/Cas9 edited samples were subjected to SDS-PAGE. ab109012 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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.
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>)
Lane 1:
Wild-type HCT116 cell lysate at 20 µg
Lane 2:
SQSTM1 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human SQSTM1 (p62) knockout HCT116 cell line (ab266871)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 55 kDa
false
- WB
Lab
Western blot - Human SQSTM1 (p62) knockout HCT116 cell line (AB266871)
Lanes 1-4 : Merged signal (red and green). Green - ab207305 observed at 55 kDa. Red - loading control ab8245 observed at 36 kDa.
ab207305 Anti-SQSTM1 / p62 antibody [EPR18351] was shown to specifically react with SQSTM1 / p62 in wild-type HCT116 cells. The band observed in CRISPR/Cas9 edited cell line ab266871 (CRISPR/Cas9 edited cell lysate ab257052) lane below 55 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and SQSTM1 / p62 CRISPR/Cas9 edited samples were subjected to SDS-PAGE. ab207305 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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.
All lanes:
Western blot - Anti-SQSTM1 / p62 antibody [EPR18351] - Autophagosome Marker (<a href='/en-us/products/primary-antibodies/sqstm1-p62-antibody-epr18351-autophagosome-marker-ab207305'>ab207305</a>)
Lane 1:
Wild-type HCT116 cell lysate at 20 µg
Lane 2:
SQSTM1 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human SQSTM1 (p62) knockout HCT116 cell line (ab266871)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 47 kDa
Observed band size: 55 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SQSTM1 (p62) knockout HCT116 cell line (AB266871)
Homozygous : 49 bp deletion in exon4
- Cell Culture
Unknown
Cell Culture - Human SQSTM1 (p62) knockout HCT116 cell line (AB266871)
Representative images of SQSTM1 knockout HCT116 cells, low and high confluency examples (top left and right respectively) and wild-type HCT116 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Human SQSTM1 (p62) knockout HCT116 cell line (AB266871)
Immunohistochemical analysis of paraffin-embedded (A) Wild-type HCT116 (human colon epithelial) cell pellet (B) SQSTM1 knockout HCT116 (ab266871) cell pellet tissue labeling SQSTM1 with ab314504 at 1/20000 (0.025 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) Wild-type HCT116 cell pellet, no staining on (B) SQSTM1 knockout HCT116 (ab266871) cell pellet. The section was incubated with ab314504 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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.
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
McCoY5a + 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
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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