HLA-E KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 8 bp deletion in exon 3.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 8 bp deletion in exon 3
EA1.2, EA2.1, HLA 6.2, HLA class I histocompatibility antigen E alpha chain, HLA class I histocompatibility antigen E alpha chain precursor, HLA class I histocompatibility antigen alpha chain E, Lymphocyte antigen, MHC, MHC HLA E alpha 1, MHC HLA E alpha 2.1, MHC class I antigen E, Major histocompatibility complex class I E, QA1
HLA-E KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 8 bp deletion in exon 3.
HEK-293T
Human
Kidney
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 8 bp deletion in exon 3
HLA-E
Knockout
CRISPR technology
Sanger Sequencing, Western blot
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). 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.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Lanes 1-3: Merged signal (red and green). Green - Anti-HLA E antibody [MEM-E/02] ab2216 observed at 40 kDa. Red - loading control Anti-beta Tubulin antibody [EP1331Y] - Microtubule Marker ab52901 observed at kDa.
Anti-HLA E antibody [MEM-E/02] ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267231 (knockout cell lysate Human HLA-E (HLA E) knockout HEK-293T cell lysate ab258454) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. Anti-HLA E antibody [MEM-E/02] ab2216 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (Anti-beta Tubulin antibody [EP1331Y] - Microtubule Marker ab52901) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-HLA E antibody [MEM-E/02] (Anti-HLA E antibody [MEM-E/02] ab2216) at 1/500 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HLA-E knockout HEK-293T cell lysate at 20 µg
Lane 3: THP-1 cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 40 kDa
Lanes 1-3: Merged signal (red and green). Green - Anti-HLA E antibody [MEM-E/02] ab2216 observed at 40 kDa. Red - loading control Anti-beta Tubulin antibody [EP1331Y] - Microtubule Marker ab52901 observed at kDa.
Anti-HLA E antibody [MEM-E/02] ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267231 (knockout cell lysate Human HLA-E (HLA E) knockout HEK-293T cell lysate ab258454) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. Anti-HLA E antibody [MEM-E/02] ab2216 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (Anti-beta Tubulin antibody [EP1331Y] - Microtubule Marker ab52901) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Representative images of HLA-E knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Allele-1: 8 bp deletion in exon3
Allele-3: 1 bp insertion in exon 3.
Allele-2: 1 bp deletion in exon 3.
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