Human CD1A knockout Jurkat cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human CD1A knockout Jurkat cell line (AB274926)
False colour image of Western blot : Anti-CD1a antibody [EP3091] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab76531 was shown to bind specifically to CD1a. A band was observed at 45-50 kDa in wild-type Jurkat cell lysates with no signal observed at this size in CD1A knockout cell line ab274926 (knockout cell lysate ab274984). To generate this image, wild-type and CD1A knockout Jurkat 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-CD1a antibody [EP3091] (<a href='/en-us/products/primary-antibodies/cd1a-antibody-ep3091-ab76531'>ab76531</a>) at 1/1000 dilution
Lane 1:
Wild-type Jurkat cell lysate at 20 µg
Lane 2:
CD1A knockout Jurkat cell lysate at 20 µg
Lane 2:
Western blot - Human CD1A knockout Jurkat cell line (ab274926)
Lane 3:
MOLT-4 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat anti-Mouse IgG H&L preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 37 kDa
Observed band size: 45-50 kDa
false
- WB
Lab
Western blot - Human CD1A knockout Jurkat cell line (AB274926)
False colour image of Western blot : Anti-CD1a antibody [EP3622] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab108309 was shown to bind specifically to CD1a. A band was observed at 45-50 kDa in wild-type Jurkat cell lysates with no signal observed at this size in CD1A knockout cell line ab274926 (knockout cell lysate ab274984). To generate this image, wild-type and CD1A knockout Jurkat 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-CD1a antibody [EP3622] (<a href='/en-us/products/primary-antibodies/cd1a-antibody-ep3622-ab108309'>ab108309</a>) at 1/1000 dilution
Lane 1:
Wild-type Jurkat cell lysate at 20 µg
Lane 2:
CD1A knockout Jurkat cell lysate at 20 µg
Lane 2:
Western blot - Human CD1A knockout Jurkat cell line (ab274926)
Lane 3:
MOLT-4 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Predicted band size: 37 kDa
Observed band size: 45-50 kDa
false
- NGS
Lab
Next Generation Sequencing - Human CD1A knockout Jurkat cell line (AB274926)
1 bp insertion after Trp30 of the WT protein
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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 2x105 cells/mL. 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 2x105 cells/mL is recommended.
- Do not allow cell density to exceed 3x106 cells/mL.
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
CD1a participates in the immune system by presenting lipid and glycolipid antigens to T lymphocytes including natural killer T (NKT) cells. It forms a complex with β2-microglobulin which is essential for its proper function and antigen presentation. CD1a expression helps differentiate between types of dendritic cells with CD1a-positive cells often engaging in specific responses to microbial lipid antigens.
Pathways
CD1a plays a significant role in the antigen presentation pathway particularly in lipid antigen presentation to T cells. This is important for initiating immune responses. It interacts with related proteins such as CD1b and CD1c which also present lipid antigens. CD1a has implications in innate and adaptive immunity and can influence pathways involved in microbial recognition and elimination.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
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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