Human STT3A knockout HEK-293T cell line
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STT3A 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: 2 bp deletion in exon 10. 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
Oligosaccharyl transferase subunit STT3A, STT3-A, STT3A, ITM1, Integral membrane protein 1, Transmembrane protein TMC, B5, TMC, Dolichyl-diphosphooligosaccharide--protein glycosyltransferase subunit STT3A
- WB
Lab
Western blot - Human STT3A knockout HEK-293T cell line (AB266320)
All lanes:
Anti-STT3A antibody at 1/200 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human STT3A knockout HEK-293T cell line (ab266320)
Lane 2:
Western blot - Human STT3A knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-stt3a-knockout-hek-293t-cell-lysate-ab259164'>ab259164</a>)
Lane 3:
Caco-2 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
false
- Sanger seq
Unknown
Sanger Sequencing - Human STT3A knockout HEK-293T cell line (AB266320)
Homozygous : 2 bp deletion in exon 10
- IP
Lab
Immunoprecipitation - Human STT3A knockout HEK-293T cell line (AB266320)
STT3A was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) whole cell lysate with ab320831 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab320831 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Blocking and diluting buffer and concentration : 5% NFDM/TBST.
In Immunoprecipitation, ab320831 was shown to bind specifically to STT3A. Target of interest was enriched at 60 kDa in wild-type HEK-293T cell lysates (lane 2) with no signal observed at this size in STT3A knockout cell line (lane 5) (lane 5, knockout cell line ab266320 / knockout cell lysate ab259164).
The bands above 60kDa are likely to be aggregation, and are absent in the KO samples. Unlike the Western Blot data, these samples have been boiled. ab181602 was used as a GAPDH loading control.
All lanes:
Immunoprecipitation - Anti-STT3A antibody [EPR29178-10] (<a href='/en-us/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a>) at 1/1000 dilution
Lane 1:
Parental HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 10 µg
Lane 2:
Parental HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3:
Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a> in Parental HEK-293T whole cell lysate
Lane 4:
STT3A KO HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 10 µg
Lane 5:
Immunoprecipitation - Human STT3A knockout HEK-293T cell line (ab266320)
Lane 6:
Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/stt3a-antibody-epr29178-10-ab320831'>ab320831</a> in STT3A KO HEK-293T whole cell lysate
Secondary
All lanes:
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution
Observed band size: 60 kDa
false
Exposure time: 84s
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Product details
We will provide viable cells that proliferate on revival.
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
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
STT3A is part of the OST complex which catalyzes the transfer of glycan from a lipid donor to asparagine residues on nascent polypeptides in a process known as co-translational N-glycosylation. This modification is essential for proper protein folding stability and activity. The OST complex with STT3A as a catalytic core is fundamental in eukaryotic cells affecting the function of numerous glycoproteins. It interacts with various components within the endoplasmic reticulum to ensure efficient glycosylation.
Pathways
The involvement of STT3A extends to the protein processing in endoplasmic reticulum pathway and is integral to the broader process of protein folding and quality control. This pathway ensures proper protein configuration and prevents the accumulation of misfolded proteins. STT3A functionally relates to proteins like calnexin and calreticulin which work in the calnexin/calreticulin cycle to assist in the folding of glycoproteins indicating its important role in maintaining protein homeostasis in cells.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
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.
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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