Human SEMA4D knockout A549 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
SEMA4D 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 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
A8, BB 18, CD 100, CD100 antigen, Coll 4, Collapsin 4, GR3, Leukocyte activation antigen CD100, M sema G, MSEMA, SEM4D_HUMAN, SEMA J, Sema 4d, Sema H, Sema domain immunoglobulin domain Ig transmembrane domain TM and short cytoplasmic domain semaphorin 4D, Semacl 2, Semaphorin C like 2, Semaphorin H, Semaphorin J, Semaphorin-4D, Semcl 2
- WB
Lab
Western blot - Human SEMA4D knockout A549 cell line (AB288912)
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 95-100 kDa in wild-type A549 cell lysates with no signal observed at this size in SEMA4D knockout cell line. To generate this image, wild-type and SEMA4D knockout A549 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='/en-us/products/primary-antibodies/semaphorin-4d-cd100-antibody-ep3569-ab134128'>ab134128</a>) at 1/2000 dilution
Lane 1:
Wild-type A549 cell lysate at 40 µg
Lane 2:
SEMA4D knockout A549 cell lysate at 40 µg
Lane 3:
HDLM-2 cell lysate at 20 µg
Lane 4:
PC-3 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
false
- NGS
Lab
Next Generation Sequencing - Human SEMA4D knockout A549 cell line (AB288912)
2 bp deletion and 62 bp deletion after Pro 208 of WT protein
Reactivity data
Product details
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.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
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.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
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.
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.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- 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.
Culture medium
F-12K + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
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)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com