Human MET (Met (c-Met)) knockout HeLa cell line
- Advanced Validation
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- WB
Lab
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Anti-MET antibody [EP1454Y] (ab51067) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab51067 was shown to bind specifically to MET. A band was observed at 40-50 kDa in in wild-type HeLa cell lysates with no signal observed at this size in MET knockout cell line. To generate this image, wild-type and MET 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (<a href='/ko/products/primary-antibodies/met-c-met-antibody-ep1454y-n-terminal-ab51067'>ab51067</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MET (Met (c-Met)) knockout HeLa cell lysate at 20 µg
Lane 3:
HT-29 cell lysate at 20 µg
Lane 4:
T-47D cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 40-50 kDa
false
- WB
Lab
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Anti-MET antibody [EPR19554-110] (ab254252) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab254252 was shown to bind specifically to MET. A band was observed at 80-140 kDa in wild-type HeLa cell lysates with no signal observed at this size in MET knockout cell line. To generate this image, wild-type and MET 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution. The full length MET protein is glycosylated and has multiple cleavage sites, so cleaved products can be detected between ~40-140 kDa
All lanes:
Western blot - Anti-Met (c-Met) antibody [EPR19554-110] (<a href='/ko/products/primary-antibodies/met-c-met-antibody-epr19554-110-ab254252'>ab254252</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
MET (Met (c-Met)) knockout HeLa cell lysate at 20 µg
Lane 3:
HT-29 cell lysate at 20 µg
Lane 4:
T-47D cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 80-140 kDa
false
- Cell Culture
Unknown
Cell Culture - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Representative images of MET knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Homozygous : 5 bp deletion in exon 2.
- WB
Lab
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Western blot : Anti-Met (c-Met) antibody [SP44] - C-terminal ab227637 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 50, 140 kDa in Wild-type HeLa cell line with no signal observed at this size in MET (C-MET) knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-Met (c-Met) antibody [SP44] - C-terminal (<a href='/ko/products/primary-antibodies/met-c-met-antibody-sp44-c-terminal-ab227637'>ab227637</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysates at 20 µg
Lane 2:
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (ab265961) at 20 µg
Lane 3:
T-47D cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 35-55 kDa,145-175 kDa
Observed band size: 50 kDa,140 kDa
false
- WB
Lab
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (AB265961)
Anti-Met (c-Met) antibody [EPR19067] (ab216574) staining at 1/1000 dilution, shown in green; Mouse GAPDH (ab8245) loading control staining at 1/20000 dilution, shown in magenta. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
Please note that we offer an alternative anti-c-MET antibody, ab227637, which has shown lower background and cleaner Western blot performance in our internal studies.
All lanes:
Western blot - Anti-Met (c-Met) antibody [EPR19067] (<a href='/ko/products/primary-antibodies/met-c-met-antibody-epr19067-ab216574'>ab216574</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human MET (Met (c-Met)) knockout HeLa cell line (ab265961) at 20 µg
Lane 3:
HT-29 cell lysate at 20 µg
Lane 4:
T-47D cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 155 kDa,121 kDa
Observed band size: 182 kDa,139 kDa
false
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 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
Zygosity
품질 관리 - STR analysis
배송 시 보관 조건
적절한 단기 보관 조건
적절한 장기 보관 조건
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.
타겟 정보
대체 명칭 보기
제품이 사용된 논문 (1)
Recent publications for all applications. Explore the 전체 목록and refine your search
eLife 12: PubMed39268701
2024
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
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