Human HLA-E (HLA E) knockout HEK-293T cell line
Human HLA-E (HLA E) knockout HEK-293T cell line
- Advanced Validation
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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, 1 bp deletion in exon 3 and 1 bp insertion in exon 3 and 8 bp deletion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
別名を表示する
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 HEK-293T cell line (AB267231)
Lanes 1-3 : Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab52901 observed at kDa.
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 ab258454) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-beta Tubulin [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 (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='/products/primary-antibodies/hla-e-antibody-mem-e-02-ab2216'>ab2216</a>) 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 2:
Western blot - Human HLA-E (HLA E) knockout HEK-293T cell line (ab267231)
Lane 3:
THP-1 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/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
- Sanger seq
Unknown
Sanger Sequencing - Human HLA-E (HLA E) knockout HEK-293T cell line (AB267231)
Allele-3 : 1 bp insertion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human HLA-E (HLA E) knockout HEK-293T cell line (AB267231)
Allele-2 : 1 bp deletion in exon 3.
- Cell Culture
Unknown
Cell Culture - Human HLA-E (HLA E) knockout HEK-293T cell line (AB267231)
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.
- Sanger seq
Unknown
Sanger Sequencing - Human HLA-E (HLA E) knockout HEK-293T cell line (AB267231)
Allele-1 : 8 bp deletion in exon3
Reactivity data
製品の詳細
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.
製品内容
出荷温度及び保存条件
遺伝子名
遺伝子編集のタイプ
遺伝子編集の方法
ノックアウト検証方法
出荷温度
短期保存温度
長期保存温度
取り扱い方法
初回取り扱いガイドライン
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.
継代培養ガイドライン
- 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.
培養培地
DMEM (High Glucose) + 10% FBS
凍結保存培地
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: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
ターゲットの情報
組合せ製品
![Flow Cytometry - Anti-HLA Class I antibody [W6/32] (AB22432)](https://content.abcam.com/products/images/hla-class-i-antibody-w6-32-ab22432--flow-cytometry-img428942.jpg)
Abcam product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com