Human OXCT1 (SCOT) knockout HeLa cell line
- Advanced Validation
- 각 태그의 뜻
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- WB
Lab
Western blot - Human OXCT1 (SCOT) knockout HeLa cell line (AB265358)
False colour image of Western blot : Anti-OXCT1/SCOT antibody staining at 0.04 μg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab224250 was shown to bind specifically to OXCT1/SCOT. A band was observed at 56 kDa in wild-type HeLa cell lysates with no signal observed at this size in OXCT1 knockout cell line ab265358 (knockout cell lysate ab258557). To generate this image, wild-type and OXCT1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-OXCT1/SCOT antibody (<a href='/ko/products/primary-antibodies/oxct1-scot-antibody-ab224250'>ab224250</a>) at 0.04 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 18 µg
Lane 2:
Western blot - Human OXCT1 (SCOT) knockout HeLa cell lysate (<a href='/ko/products/cell-lysates/human-oxct1-scot-knockout-hela-cell-lysate-ab258557'>ab258557</a>) at 18 µg
Lane 3:
Jurkat cell lysate at 18 µg
Lane 4:
A549 cell lysate at 18 µg
Secondary
Lanes 1 - 4:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 4:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDa,37 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human OXCT1 (SCOT) knockout HeLa cell line (AB265358)
Allele-2 : 1 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human OXCT1 (SCOT) knockout HeLa cell line (AB265358)
Allele-1 : 13 bp deletion in exon 1.
보완 제품
Reactivity 정보
제품 세부 정보
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.
제품 구성
특성 및 보관 정보
유전자명
유전자 편집 종류
유전자 편집 방식
Knockout 검증
배송 시 보관 조건
적절한 단기 보관 조건
적절한 장기 보관 조건
추가 정보
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
OXCT1 functions as an important enzyme in ketolysis which is the breakdown of ketone bodies for energy production. It does not operate within a large complex but interacts closely with other enzymes involved in similar metabolic processes. OXCT1 ensures that tissues efficiently convert ketone bodies like acetoacetate and beta-hydroxybutyrate into acetyl-CoA which then feeds into the citric acid cycle to fulfill energy requirements especially when carbohydrates are sparse.
Pathways
OXCT1 is an important component of the ketone body metabolism pathway linking it to energy homeostasis and ketogenesis. It directly interacts with enzymes like acetoacetyl-CoA thiolase and beta-hydroxybutyrate dehydrogenase. Through these associations it integrates into larger metabolic pathways such as the citric acid cycle which is essential for energy production under conditions when lipids become a primary source for energy generation.
품질 관리
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
초기 취급 지침
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 가이드라인
- 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.
배양 배지
DMEM (High Glucose) + 10% FBS
동결보존 배지
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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