Human CD74 knockout Raji cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
Western blot - Human CD74 knockout Raji cell line (AB273876)
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 (<a href='/en-us/products/cell-lysates/human-cd74-knockout-raji-cell-lysate-ab273830'>ab273830</a>) at 20 µg
Lane 3:
Daudi 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
- WB
Lab
Western blot - Human CD74 knockout Raji cell line (AB273876)
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 ° 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 (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
Predicted band size: 34 kDa
Observed band size: 35 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human CD74 knockout Raji cell line (AB273876)
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift : 100%
- NGS
Supplier Data
Next Generation Sequencing - Human CD74 knockout Raji cell line (AB273876)
1 bp deletion after Arg93 of the WT protein
Complementary Products
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 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.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 4x105 cells/mL is recommended.
- A maximum of 3x106 viable cells/mL is obtainable.
Culture medium
RPMI + 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
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
Target data
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