Human CSF1 knockout U-87 MG cell line
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- WB
Lab
Western blot - Human CSF1 knockout U-87 MG cell line (AB288711)
Western blot : Rabbit monoclonal [EPR20948] to M-CSF ab233387 staining at 1/1000 dilution, shown in black, total protein stain shown in pink. A band indicated the mature CSF1 was observed at 44 kDa in Wild type U-87 MG Supernatant cell lysates with no signal observed at this size in CSF1 knockout U-87 MG supernatant cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 HRP (H+L) at 1/20,000 dilution.
All lanes:
Western blot - Anti-M-CSF antibody [EPR20948] (<a href='/en-us/products/primary-antibodies/m-csf-antibody-epr20948-ab233387'>ab233387</a>) at 1/1000 dilution
Lane 1:
Wild type U-87 MG Supernatant at 15 µL
Lane 2:
Western blot - Human CSF1 knockout U-87 MG cell line (ab288711) at 15 µL
Lane 3:
THP-1 supernatant at 15 µL
Lane 4:
A549 supernatant at 15 µL
Lane 5:
T-47D supernatant at 15 µL
Secondary
All lanes:
Goat anti-Rabbit HRP (H+L) at 1/20000 dilution
true
Exposure time: 10s
- Sanger seq
Supplier Data
Sanger Sequencing - Human CSF1 knockout U-87 MG cell line (AB288711)
67 bp deletion in exon 4.
Complementary Products
Product details
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
Zygosity
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.
Culture medium
EMEM + 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
M-CSF influences the production differentiation and function of macrophages and osteoclasts. It is not part of a large complex but interacts with its receptor the CSF1R (colony-stimulating factor 1 receptor) on the surface of target cells. This interaction promotes the survival and proliferation of the target cells playing significant roles in immune response and bone homeostasis by regulating osteoclast development.
Pathways
The interaction of M-CSF with its receptor is central to several biological pathways notably the immune system pathway and bone resorption pathway. Within these pathways the binding of M-CSF to CSF1R activates downstream signaling cascades such as the PI3K/AKT and MAPK pathways importantly affecting cell survival and differentiation. The M-CSF pathway intersects with other cytokines and factors like GM-CSF (granulocyte-macrophage colony-stimulating factor) which also regulate immune cell dynamics.
Quality control
STR analysis
VWA, CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
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