Human CD1A knockout Jurkat cell line (ab274926)
Overview
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Product name
Human CD1A knockout Jurkat cell line -
Parental Cell Line
Jurkat -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 99% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Tested applications
Suitable for: WBmore details -
Biosafety level
1 -
General notes
Recommended control: 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.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: RPMI + 10% FBS
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 1x105 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 1x105 cells/mL is recommended.
- Do not allow the cell density to exceed 3x106.
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.
We will provide viable cells that proliferate on revival.
Properties
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Number of cells
1 x 106 cells/vial, 1 mL -
Adherent /Suspension
Suspension -
Tissue
Blood -
Cell type
T cell lymphoblast-like -
Disease
Non-Hodgkin Lymphoma -
Gender
Male -
Mycoplasma free
Yes -
Storage instructions
Shipped on Dry Ice. Store in liquid nitrogen. -
Storage buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether -
Research areas
Target
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Function
Antigen-presenting protein that binds self and non-self lipid and glycolipid antigens and presents them to T-cell receptors on natural killer T-cells. -
Tissue specificity
Expressed on cortical thymocytes, epidermal Langerhans cells, dendritic cells, on certain T-cell leukemias, and in various other tissues. -
Sequence similarities
Contains 1 Ig-like (immunoglobulin-like) domain. -
Cellular localization
Cell membrane. Endosome membrane. Subject to intracellular trafficking between the cell membrane and endosomes. Localizes to cell surface lipid rafts. - Information by UniProt
Associated products
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab274926 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration.
|
Notes |
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WB
Use at an assay dependent concentration. |
Images
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1 bp insertion after Trp30 of the WT protein
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All lanes : Anti-CD1a antibody [EP3622] (ab108309) at 1/1000 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : CD1A knockout Jurkat cell lysate
Lane 3 : MOLT-4 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 45-50 kDa why is the actual band size different from the predicted?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.
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All lanes : Anti-CD1a antibody [EP3091] (ab76531) at 1/1000 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : CD1A knockout Jurkat cell lysate
Lane 3 : MOLT-4 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 45-50 kDa why is the actual band size different from the predicted?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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab274926 has not yet been referenced specifically in any publications.