Human ACO1 knockout U-2 OS cell line
- Advanced Validation
- 각 태그의 뜻
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ACO1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
대체 명칭 보기
Iron-responsive element-binding protein 1, Iron regulatory protein 1, ACO1, IREB1, Cytoplasmic aconitate hydratase, IRP1, Citrate hydro-lyase, IRE-BP 1, Ferritin repressor protein, Aconitase
- WB
Lab
Western blot - Human ACO1 knockout U-2 OS cell line (AB261884)
Lanes 1 - 4 : Merged signal (red and green). Green - ab183721 observed at 98 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab183721 was shown to specifically react with ACO1 in wild-type U-2 OS cells as signal was lost in ACO1 knockout cell line ab261884 (knockout cell lysate ab261693). Wild-type and ACO1 knockout samples were subjected to SDS-PAGE. ab183721 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Aconitase 1/ACO1 antibody [EPR7226(2)] (<a href='/ko/products/primary-antibodies/aconitase-1-aco1-antibody-epr72262-ab183721'>ab183721</a>) at 1 µg/mL
Lane 1:
Wild-type U20S whole cell lysate at 20 µg
Lane 2:
ACO1 knockout U-2 OS whole cell lysate at 20 µg
Lane 2:
Western blot - Human ACO1 knockout U-2 OS cell line (ab261884)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Predicted band size: 98 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human ACO1 knockout U-2 OS cell line (AB261884)
2 bp insertion after Thr257 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human ACO1 knockout U-2 OS cell line (AB261884)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 100%
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
ACO1 connects energy production and iron regulation. It binds to iron-responsive elements when the iron is scarce and acts in its aconitase form when iron levels are sufficient. ACO1 is not part of a larger protein complex but plays an important regulatory role in balancing cellular energy output and iron homeostasis. It affects both glycolysis and the electron transport chain indirectly by altering the availability of citric acid cycle intermediates which are important for ATP production.
Pathways
ACO1 integrates into both the citric acid cycle and the iron regulatory pathway. The citric acid cycle also called the Krebs cycle is fundamental for ATP production and ACO1 serves as a pivotal point within this cycle. Moreover through its iron-regulatory function it connects to iron metabolism pathways that are vital for various cellular processes. ACO1 activity also influences other proteins like ferritin by regulating iron storage and transport mechanisms through its dual functionality.
품질 관리
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
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
McCoY5a + 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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