Human HLA-E (HLA E) knockout A549 cell line
- Advanced Validation
- What is this?
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HLA-E KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 2. 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
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
- WB
Lab
Western blot - Human HLA-E (HLA E) knockout A549 cell line (AB267081)
Lanes 1-4 : Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab181602 observed at 36 kDa.
ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267081 (knockout cell lysate ab258453) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and 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] (<a href='/en-us/products/primary-antibodies/hla-e-antibody-mem-e-02-ab2216'>ab2216</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
HLA-E knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human HLA-E (HLA E) knockout A549 cell line (ab267081)
Lane 3:
THP-1 cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa
false
- Cell Culture
Unknown
Cell Culture - Human HLA-E (HLA E) knockout A549 cell line (AB267081)
Representative images of HLA-E knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human HLA-E (HLA E) knockout A549 cell line (AB267081)
Homozygous : 5 bp deletion in exon2
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
Zygosity
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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