HLA-A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 17 bp deletion Frameshift = 99.8%.
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
HLA-A KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 17 bp deletion Frameshift = 99.8%.
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.
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.
HLA-A also known as Human Leukocyte Antigen A is a protein that is part of the major histocompatibility complex (MHC) class I molecules. It has an approximate molecular mass of 44 kDa. HLA-A is expressed on the surface of nearly all nucleated cells and interacts with CD8+ T lymphocytes. Known for its role in immune recognition HLA-A presents endogenous peptide antigens to the T-cell receptor on cytotoxic T cells initiating immune responses to eliminate infected or malignant cells.
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.
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.
HLA-A has been associated with autoimmune diseases and transplant rejection. It plays a significant role in conditions like ankylosing spondylitis where specific HLA-A subtypes contribute to disease susceptibility. Additionally mismatches in HLA-A typing can lead to transplant rejection. In such scenarios anti-HLA antibodies can develop causing damage to transplant tissues. The complex interaction of HLA-A with related proteins such as other MHC class I molecules influences immune response outcomes in these diseases.
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 - ab86597 observed at 40 kDa. Red - loading control, Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker 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 Human HLA-A knockout A-431 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 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker 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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed 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] (ab86597) 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
Performed under reducing conditions.
Lanes 1 - 4: Merged signal (red and green). Green - Anti-HLA A antibody [EP1395Y] ab52922 observed at 40 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
Anti-HLA A antibody [EP1395Y] 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 Human HLA-A knockout A-431 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 Anti-HLA A antibody [EP1395Y] ab52922 and Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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] (Anti-HLA A antibody [EP1395Y] ab52922) 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
Performed under reducing conditions.
X = 17 bp deletion
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