Human CD58 knockout Raji cell line
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- NGS
Lab
Next Generation Sequencing - Human CD58 knockout Raji cell line (AB290422)
107 bp deletion in exon 3, CCDS888.1
- WB
Lab
Western blot - Human CD58 knockout Raji cell line (AB290422)
Western blot : Anti-CD58 antibody [EPR24012-147] ab275392 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 55-70 kDa in Wild-type Raji cell lysates with no signal observed at this size in CD58 knockout Raji 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-CD58 antibody [EPR24012-147] (<a href='/en-us/products/primary-antibodies/cd58-antibody-epr24012-147-ab275392'>ab275392</a>) at 1/1000 dilution
Lane 1:
Wild-type Raji at 20 µg
Lane 2:
Western blot - Human CD58 knockout Raji cell line (ab290422) at 20 µg
Lane 3:
Wild-type HeLa at 20 µg
Lane 4:
CD58 knockout HeLa cells at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 28 kDa,52-60 kDa
Observed band size: 55-70 kDa,37 kDa
false
Complementary Products
![Flow Cytometry - Anti-CD58 antibody [EPR24012-147] (AB275392)](https://content.abcam.com/products/images/cd58-antibody-epr24012-147-ab275392--flow-cytometry-img71382.jpg)
Reactivity data
Product details
Recommended control: Human wild-type Raji cell line (ab290717). 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.
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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 - 5x105 cells/mL(for initial passages it is recomended to culture the cells in the higher range of recomended seeding density). Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 4x105 cells/mL is recommended.
- A maximum of 3x106 viable cells/mL is obtainable.
Culture medium
RPMI + 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
Lymphocyte function-associated antigen 3 acts as a significant participant in the immune system. It plays a role in T cell activation and facilitates interactions between T cells and antigen-presenting cells. CD58 forms part of a complex network of cell surface proteins that enhances immune surveillance and response. Its role highlights its importance in adaptive immunity by promoting cell-to-cell communication and activation.
Pathways
CD58 participates in the cell adhesion pathways. It interacts closely with CD2 which is involved in the activation and signaling of T cells. Through this interaction CD58 plays roles in pathways like T cell receptor signaling and immune response. It enhances cell-cell adhesion helping T cells maintain their position during immune surveillance and activation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Suspension
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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