Human CD74 knockout Raji cell lysate
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CD74 KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion Frameshift: 100%.
View Alternative Names
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
- WB
Lab
Western blot - Human CD74 knockout Raji cell lysate (AB273830)
Lane 1 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
Lane 2 : CD74 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
Lane 3 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate (20 ug)
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)
Lanes 1 - 4 : Merged signal (red and green). Green - ab64772 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
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 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 ab64772 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 h at room temperature before imaging.
All lanes:
Western blot - Anti-CD74 antibody (<a href='/en-us/products/primary-antibodies/cd74-antibody-ab64772'>ab64772</a>) 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 (<a href='/en-us/products/cell-lines/human-cd74-knockout-raji-cell-line-ab273876'>ab273876</a>)
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
Predicted band size: 34 kDa
Observed band size: 35 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human CD74 knockout Raji cell lysate (AB273830)
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift : 100%
- WB
Lab
Western blot - Human CD74 knockout Raji cell lysate (AB273830)
Lanes 1 - 4 : Merged signal (red and green). Green - ab108393 observed at 34 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
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 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 ab108393 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-CD74 antibody [EPR4064] (<a href='/en-us/products/primary-antibodies/cd74-antibody-epr4064-ab108393'>ab108393</a>) at 1/1000 dilution
Lane 1:
Wild-type Raji cell lysate at 20 µg
Lane 2:
Western blot - Human CD74 knockout Raji cell lysate (ab273830) at 20 µg
Lane 3:
Daudi cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 4:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDa,55 kDa
false
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
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.
Pathways
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Suspension
Gender
Male
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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