Human HLA-A knockout A-431 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
HLA-A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 17 bp deletion Frameshift = 99.8%. 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
Antigen Presenting Molecule, HLA class I histocompatibility antigen, A 1 alpha chain, Leukocyte antigen class I A, MHC class I antigen HLA A heavy chain, Major histocompatibility complex, class I, A
- WB
Supplier Data
Western blot - Human HLA-A knockout A-431 cell line (AB261894)
Lanes 1 - 4 : Merged signal (red and green). Green - ab52922 observed at 40 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab52922 was shown to react with HLA-A in wild-type A-431 cells in Western blot Loss of signal was observed when HLA-A knockout cell line ab261894 (knockout cell lysate ab261703) was used. Wild-type A-431 and HLA-A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab52922 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-HLA A antibody [EP1395Y] (<a href='/en-us/products/primary-antibodies/hla-a-antibody-ep1395y-ab52922'>ab52922</a>) at 1/10000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
HLA A knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human HLA-A knockout A-431 cell line (ab261894)
Lane 3:
A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
false
- WB
Lab
Western blot - Human HLA-A knockout A-431 cell line (AB261894)
Lanes 1 - 3 : Merged signal (red and green). Green - ab86597 observed at 40 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab86597 was shown to react with HLA-A in wild-type A-431 cells in Western blot Loss of signal was observed when HLA-A knockout cell line ab261894 (knockout cell lysate ab261703) was used. Wild-type A-431 and HLA-A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab86597 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Anti-HLA Class I A3 antibody [7g7h8] (<a href='/en-us/products/unavailable/hla-class-i-a3-antibody-7g7h8-ab86597'>ab86597</a>) at 5 µg/mL
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2:
HLA-A knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2:
Western blot - Human HLA-A knockout A-431 cell line (ab261894)
Lane 3:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
false
- NGS
Lab
Next Generation Sequencing - Human HLA-A knockout A-431 cell line (AB261894)
X = 17 bp deletion
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
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.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
This target functions as a component of the MHC class I molecule complex. HLA-A binds peptides derived from proteasomal degradation of intracellular proteins and presents these peptides on the cell surface. The function plays an essential role in immune surveillance and distinction between self and non-self. This antigen presentation is important for immune responses against viral infections and tumor surveillance influencing the activation and proliferation of T lymphocytes.
Pathways
HLA-A plays a significant role in the antigen processing and presentation pathway. It is closely related to the TAP (Transporter associated with Antigen Processing) proteins which transport antigenic peptides into the endoplasmic reticulum for loading onto MHC class I molecules. Additionally the target interacts with the CD8 molecule on cytotoxic T cells. This interaction is important within the immune response pathway contributing to the recognition and destruction of infected cells.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Immunoprecipitation - Anti-HLA A antibody [EP1395Y] (AB52922)](https://content.abcam.com/products/images/hla-a-antibody-ep1395y-ab52922--immunoprecipitation-img273.jpg)
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