CD74 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 = 1 bp deletion Frameshift: 100%.
CD74 antigen, CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated), CD74 molecule, CD74 molecule, major histocompatibility complex, class II invariant chain, DHLAG, Gamma chain of class II antigens, HG2A_HUMAN, HLA class II histocompatibility antigen gamma chain, HLA-DR antigens-associated invariant chain, HLA-DR-gamma, HLADG, Ia antigen-associated invariant chain, Ia associated invariant chain, Ia gamma, Ii, Invariant polypeptide of major histocompatibility complex class II antigen associated, MHC HLA-DR gamma chain, Major histocompatibility complex class II invariant chain, P35, Protein 41, la-gamma, p33
CD74 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 = 1 bp deletion Frameshift: 100%.
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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 - 5x105 cells/mL(for initial passages it is recomended to culture the cells in the higher range of recomended seeding density). Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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.
CD74 also known as the invariant chain or li is a protein with a mass of about 33 kDa. It is widely expressed on the surface of immune cells such as B cells dendritic cells and macrophages. CD74 plays an important role in the immune system by acting as a chaperone for major histocompatibility complex class II (MHC II) molecules guiding their migration to endosomes where they encounter antigenic peptides. The CD74 protein also functions as a cell surface receptor for macrophage migration inhibitory factor (MIF) enhancing the immune response process.
CD74 facilitates multiple immune regulatory processes. It forms a complex with the MHC II molecules aiding in their proper folding and peptide loading in the endoplasmic reticulum. This complex then travels through the Golgi apparatus towards the lysosomal compartments where antigens are presented to initiate adaptive immune responses. In addition CD74 is involved in signal transduction pathways that regulate cell proliferation and survival influenced by interactions with MIF and other molecules.
The role of CD74 extends into both the antigen presentation and MIF signaling pathways. Within the antigen presentation pathway CD74's activity is tightly connected with MHC II and subsequently to CD4+ T cells facilitating the activation of these immune cells. The MIF signaling pathway involves CD74 interacting with CD44 a receptor that further propagates the signaling cascades essential for immune modulation and cell survival.
CD74 is linked to inflammatory and autoimmune conditions such as rheumatoid arthritis and systemic lupus erythematosus. Aberrant expression of CD74 influences the presentation of autoantigens and may contribute to the chronic inflammation seen in these disorders. Its connection to MIF which is a pro-inflammatory cytokine further implicates CD74 in these autoimmune scenarios by promoting persistent immune activation. Researchers are also exploring CD74 as a therapeutic target in certain cancers due to its influence on immune surveillance and tumor progression.
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Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift: 100%
Anti-CD74 antibody [EPR4064] ab108393 was shown to react with CD74 in wild-type Raji cells in Western blot with loss of signal observed in CD74 knockout cell line ab273876 (knockout cell lysate Human CD74 knockout Raji cell lysate ab273830). Wild-type Raji and CD74 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-CD74 antibody [EPR4064] ab108393 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 h at room temperature before imaging.
All lanes: Western blot - Anti-CD74 antibody [EPR4064] (Anti-CD74 antibody [EPR4064] ab108393) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: CD74 knockout Raji cell lysate at 20 µg
Lane 2: Western blot - Human CD74 knockout Raji cell line (ab273876)
Lane 3: Daudi cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Anti-CD74 antibody ab64772 was shown to react with CD74 in wild-type Raji cells in Western blot with loss of signal observed in CD74 knockout cell line ab273876 (knockout cell lysate Human CD74 knockout Raji cell lysate ab273830). Wild-type Raji and CD74 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-CD74 antibody ab64772 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 h at room temperature before imaging.
All lanes: Western blot - Anti-CD74 antibody (Anti-CD74 antibody ab64772) at 1 µg/mL
Lane 1: Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 2: CD74 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human CD74 knockout Raji cell line (ab273876)
Lane 3: Daudi (Human Burkitt's lymphoma 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.
Predicted band size: 34 kDa
Observed band size: 35 kDa
1 bp deletion after Arg93 of the WT protein
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