Human BRCA2 knockout (hetero) MCF7 cell line
- Advanced Validation
- 각 태그의 뜻
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BRCA2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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
Breast cancer type 2 susceptibility protein, FANCD1, Fanconi anemia group D1 protein, FACD, BRCA2
- WB
Lab
Western blot - Human BRCA2 knockout (hetero) MCF7 cell line (AB286285)
Anti-BRCA2 antibody [EPR23442-43] (ab239375) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab239375 was shown to bind specifically to BRCA2. A band was observed at 290 kDa in wild-type MCF7 cell lysates with no signal observed at this size in BRCA2 knockout cell line ab286285 (knockout cell lysate ab300210). To generate this image, wild-type and BRCA2 knockout MCF7 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
All lanes:
Western blot - Anti-BRCA2 antibody [EPR23442-43] (<a href='/ko/products/primary-antibodies/brca2-antibody-epr23442-43-ab239375'>ab239375</a>) at 1/500 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
Western blot - Human BRCA2 knockout MCF7 cell lysate (ab300210) at 20 µg
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
PC-3 cell lysate at 20 µg
Lane 5:
HT-29 cell lysate at 20 µg
Secondary
Lanes 1 - 5:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 5:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 290 kDa
false
- NGS
Lab
Next Generation Sequencing - Human BRCA2 knockout (hetero) MCF7 cell line (AB286285)
193 bp deletion after the Val1497 of the WT protein
보완 제품
Reactivity 정보
제품 세부 정보
Recommended control: Human wild-type MCF7 cell line (ab288560). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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.
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
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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
배양 배지
MEM + 10% FBS + 0.01 mg/ml bovine insulin
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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