Human SELENBP1 (Selenium Binding Protein 1/SBP) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Unknown
Western blot - Human SELENBP1 (Selenium Binding Protein 1/SBP) knockout HeLa cell line (AB265279)
Lanes 1-3 : Merged signal (red and green). Green - ab90135 observed at 60 kDa. Red - loading control ab8245 observed at 36 kDa.
ab90135 Anti-Selenium Binding Protein 1/SBP antibody was shown to specifically react with Selenium Binding Protein 1/SBP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265279 (knockout cell lysate ab257662) was used. Wild-type and Selenium Binding Protein 1/SBP knockout samples were subjected to SDS-PAGE. ab90135 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-Selenium Binding Protein 1/SBP antibody (<a href='/en-us/products/primary-antibodies/selenium-binding-protein-1-sbp-antibody-ab90135'>ab90135</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
SELENBP1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SELENBP1 (Selenium Binding Protein 1/SBP) knockout HeLa cell line (ab265279)
Lane 3:
HT-29 cell lysate at 20 µg
Lane 4:
SW 480 cell lysate at 20 µg
Secondary
Lanes 1 - 4:
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
Lane 4:
<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: 52 kDa
Observed band size: 60 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SELENBP1 (Selenium Binding Protein 1/SBP) knockout HeLa cell line (AB265279)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human SELENBP1 (Selenium Binding Protein 1/SBP) knockout HeLa cell line (AB265279)
Allele-1 : 11 bp deletion in exon 2.
Complementary Products
Cell Lines & Lysates
AB257662
Human SELENBP1 (Selenium Binding Protein 1/SBP) knockout HeLa cell lysate
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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
Selenium Binding Protein 1 interacts with selenium implicating it in various metabolic processes. It does not act as part of a larger complex but functions independently to influence cellular responses to selenium levels. Research shows that SBP1 plays a role in antioxidant defense and cellular detoxification suggesting a protective function against oxidative stress. This regulation supports various metabolic pathways by modulating selenium-dependent enzyme activity.
Pathways
Selenium Binding Protein 1 has been connected to the antioxidant defense pathway and the detoxification process. It interacts with proteins involved in these pathways such as glutathione peroxidases essential for eliminating oxidative damage in cells. By influencing these pathways SBP1 helps maintain cellular homeostasis highlighting its role in the broader metabolic network.
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'.
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