Human CD58 knockout HeLa cell line
- Advanced Validation
- What is this?
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CD58 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 1 and 2 bp deletion in exon 1. 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
AG3, CD58 antigen, CD58 antigen (lymphocyte function associated antigen 3)1, CD58 antigen, (lymphocyte function associated antigen 3), CD58 molecule, FLJ23181, FLJ43722, LFA3_HUMAN, Lymphocyte Function Associated Antigen Type 3, Lymphocyte function-associated antigen 3, Surface glycoprotein LFA-3
- WB
Lab
Western blot - Human CD58 knockout HeLa cell line (AB265947)
False colour image of Western blot : Anti-CD58 antibody [EPR24012-147] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab275392 was shown to bind specifically to CD58. A band was observed at 55 kDa in wild-type HeLa cell lysates with no signal observed at this size in CD58 knockout cell line ab265947 (knockout cell lysate ab257880). To generate this image wild-type and CD58 knockout HeLa 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 °. 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 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 HeLa cell lysate at 20 µg
Lane 2:
CD58 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CD58 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-cd58-knockout-hela-cell-lysate-ab257880'>ab257880</a>)
Lane 2:
Western blot - Human CD58 knockout HeLa cell line (ab265947)
Lane 3:
Raji cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 28 kDa
Observed band size: 55 kDa
false
- WB
Lab
Western blot - Human CD58 knockout HeLa cell line (AB265947)
Lanes 1-4 : Merged signal (red and green). Green - ab196648 observed at 43 kDa. Red - loading control ab8245 observed at 37 kDa.
ab196648 Anti-CD58 antibody [EP15041] was shown to specifically react with CD58 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265947 (knockout cell lysate ab257880) was used. Wild-type and CD58 knockout samples were subjected to SDS-PAGE. ab196648 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-CD58 antibody [EP15041] (<a href='/en-us/products/primary-antibodies/cd58-antibody-ep15041-ab196648'>ab196648</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CD58 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CD58 knockout HeLa cell line (ab265947)
Lane 3:
HAP1 cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 28 kDa
Observed band size: 43 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CD58 knockout HeLa cell line (AB265947)
Allele-1 : 11 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human CD58 knockout HeLa cell line (AB265947)
Allele-2 : 2 bp deletion in exon 1.
Reactivity data
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
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
DMEM (High Glucose) + 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
Adherent
Gender
Female
Target data
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)
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