Human SPP1 knockout A549 cell line
- Advanced Validation
- 각 태그의 뜻
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SPP1 KO cell line available to order. KO validated by Next Generation Sequencing, 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
OPN, BNSP, Urinary stone protein, SPP1, PSEC0156, Uropontin, Nephropontin, SPP-1, Osteopontin, Bone sialoprotein 1, Secreted phosphoprotein 1
- NGS
Lab
Next Generation Sequencing - Human SPP1 knockout A549 cell line (AB287778)
1 bp insertion after Pro75
- WB
Lab
Western blot - Human SPP1 knockout A549 cell line (AB287778)
Western blot : Anti-Osteopontin antibody [EPR21139-316] ab214050 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 51 kDa. 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-Osteopontin antibody [EPR21139-316] (<a href='/ko/products/primary-antibodies/osteopontin-antibody-epr21139-316-ab214050'>ab214050</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 BFA (5 ug/mL, 6h) cell lysates at 20 µg
Lane 2:
Western blot - Human SPP1 knockout A549 cell line (ab287778) at 20 µg
Lane 3:
SPP1 knockout A549 ab287778 BFA (5 ug/mL, 6h) cell lysates at 20 µg
Lane 4:
SPP1 knockout A549 ab287778 BFA (0 ug/mL, 6h) cell lysates at 20 µg
Lane 5:
HepG2 BFA (5 ug/mL, 6h) cell lysates at 20 µg
Lane 6:
HepG2 BFA (0 ug/mL, 6h) cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 51 kDa
false
Reactivity 정보
제품 세부 정보
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (ab288558). 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
추가 정보
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Osteopontin affects cell adhesion migration and survival. It does not form part of a known large complex but interacts closely with its integrin receptors. Osteopontin modulates immune responses and aids in bone remodeling by affecting osteoclast and osteoblast functions. It is also influential in inflammation processes where it orchestrates the recruitment and activation of macrophages. The osteopontin function is significant in both physiological processes and pathologies marking it as an important protein to study in numerous biological contexts.
Pathways
Osteopontin is integral to both the bone remodeling and immune response pathways. It collaborates with proteins like osteocalcin and matrix metalloproteinases within these processes. In the bone remodeling pathway osteopontin mediates communication between osteoclasts and osteoblasts impacting bone resorption and formation. In the immune response pathway it coordinates with cytokines such as interleukin-6 to modulate immune cell activities. Its roles in these pathways indicate its multifunctional nature and its impact across different biological systems.
품질 관리
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
배양 배지
F-12K + 10% FBS
동결보존 배지
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
제품이 사용된 논문 (2)
Recent publications for all applications. Explore the 전체 목록 and refine your search
International journal of molecular sciences 26: PubMed40943512
2025
Applications
Unspecified application
Species
Unspecified reactive species
Clinical and experimental medicine 25:55 PubMed39937284
2025
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
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