Human SEMA4D knockout HCT116 cell line
- Advanced Validation
- 각 태그의 뜻
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- WB
Lab
Western blot - Human SEMA4D knockout HCT116 cell line (AB289144)
Western blot : Anti-SEMA4D antibody [30/CD100] (ab307685) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab307685 was shown to bind specifically to SEMA4D. A band was observed at 90-120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in SEMA4D knockout cell line. To generate this image, wild-type and SEMA4D knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Semaphorin 4D/CD100 antibody [30/CD100] (<a href='/ko/products/primary-antibodies/semaphorin-4d-cd100-antibody-30-cd100-ab307685'>ab307685</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human SEMA4D knockout HCT116 cell line (ab289144)
Lane 2:
SEMA4D knockout HCT 116 cell lysate at 20 µg
Lane 3:
HDLM-2 UNBOILED cell lysate at 20 µg
Lane 4:
PC-3 UNBOILED cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Observed band size: 90-120 kDa
false
- WB
Lab
Western blot - Human SEMA4D knockout HCT116 cell line (AB289144)
Western blot : Anti-SEMA4D antibody [EP3569] (ab134128) staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab134128 was shown to bind specifically to SEMA4D. A band was observed at 90-120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in SEMA4D knockout cell line. To generate this image, wild-type and SEMA4D knockout HCT 116 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-Semaphorin 4D/CD100 antibody [EP3569] (<a href='/ko/products/primary-antibodies/semaphorin-4d-cd100-antibody-ep3569-ab134128'>ab134128</a>) at 1/2000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human SEMA4D knockout HCT116 cell line (ab289144)
Lane 2:
SEMA4D knockout HCT 116 cell lysate at 20 µg
Lane 3:
HDLM-2 UNBOILED cell lysate at 20 µg
Lane 4:
PC-3 UNBOILED cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 90-120 kDa
false
- NGS
Lab
Next Generation Sequencing - Human SEMA4D knockout HCT116 cell line (AB289144)
86 bp deletion in exon 7, CCDS6685.1
보완 제품
Primary Antibodies
AB134128
Anti-Semaphorin 4D/CD100 antibody [EP3569]
KO Validated|RabMAb|Recombinant
![Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 4D/CD100 antibody [EP3569] (AB134128)](https://content.abcam.com/products/images/semaphorin-4d-cd100-antibody-ep3569-ab134128--immunocytochemistry-immunofluorescence-img105434.jpg)
- 0
- 0
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ahsp antibody [EPR12319] (AB180861)](https://content.abcam.com/products/images/ahsp-antibody-epr12319-ab180861--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img122752.jpg)
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
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
CD100 influences various cellular functions by interacting with multiple receptors such as Plexin-B1 and CD72 as part of a larger signaling complex. It contributes to immune response regulation by promoting T-cell activation and B-cell aggregation which are essential for adaptive immunity. In the nervous system it guides neural development including axon growth and synaptic plasticity. CD100's interaction with Plexin-B1 plays a significant role in these developmental processes often affecting cell morphology and movement.
Pathways
Semaphorin 4D is involved in both immune and neural signaling pathways. In the immune system it influences the PI3K-Akt signaling pathway which impacts cell survival and immune cell activation. It associates with pathways involving Plexin-B1 and RhoA to regulate cytoskeletal dynamics affecting cell motility and growth. Additionally CD100's role in neural pathways intersects with proteins like neuropilin-1 influencing semaphorin-mediated signaling essential for neural connectivity.
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.
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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