CD1A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 99%.
Images
Key facts
- Cell type
Jurkat
- Species or organism
Human
- Tissue
Blood
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
- Mutation description
Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 99%
Alternative names
CD1, CD1A Antigen, CD1A antigen, a polypeptide, CD1A_HUMAN, CD1a molecule, Cortical thymocyte antigen CD1A, Differentiation antigen CD1 alpha 3, FCB 6, HTA 1, OTTHUMP00000018907, R 4, T 6, T-cell surface antigen T6/Leu-6, T-cell surface glycoprotein CD1a, cluster of differentiation 1 A, epidermal dendritic cell marker CD1a, hTa1 thymocyte antigen
Recommended products
CD1A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 99%.
Key facts
- Cell type
Jurkat
- Species or organism
Human
- Tissue
Blood
- Form
Liquid
- Knockout validation
Next Generation Sequencing, Western blot
- Mutation description
Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 99%
- Disease
Non-Hodgkin Lymphoma
- Concentration
Properties
- Gene name
CD1A
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Next Generation Sequencing, Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 1 US: 1
- Adherent/suspension
Suspension
- Gender
Male
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.
Storage
- Shipped at conditions
Dry Ice
- Appropriate short-term storage conditions
-196°C
- Appropriate long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type Jurkat cell line (Human wild-type Jurkat cell line ab271143). 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, 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.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
CD1a also known as T6 antigen or CD1 is a protein with an estimated molecular mass of 37 kDa. It is expressed mainly on the surface of Langerhans cells in the skin and dendritic cells in the thymus. CD1a belongs to the CD1 family which presents lipid antigens to T cells. The protein is an important marker in immunophenotyping of these dendritic cells making it useful in flow cytometry applications.
- 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.
- Associated diseases and disorders
CD1a is associated with certain skin conditions like Langerhans Cell Histiocytosis and psoriasis. It is involved in the pathogenesis of these diseases by affecting immune responses in the skin. In Langerhans Cell Histiocytosis CD1a-positive cells accumulate abnormally. Additionally CD1a is often studied alongside related proteins like CD207 (Langerin) to understand these conditions better.
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3 product images
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] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD1a antibody [EP3622] 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 Human CD1A knockout Jurkat 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-CD1a antibody [EP3622] (Anti-CD1a antibody [EP3622] ab108309) 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 3: MOLT-4 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 45-50 kDa
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] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD1a antibody [EP3091] 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 Human CD1A knockout Jurkat 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-CD1a antibody [EP3091] (Anti-CD1a antibody [EP3091] ab76531) 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 3: MOLT-4 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 45-50 kDa
Next Generation Sequencing - Human CD1A knockout Jurkat cell line (ab274926)
1 bp insertion after Trp30 of the WT protein
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