Human CD27A knockout Raji cell line
- Advanced Validation
- What is this?
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CD27 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 = 10 bp deletion Frameshift: 100%. 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
CD27 antigen, CD27 molecule, CD27L receptor, CD27_HUMAN, LPFS2, MGC20393, OTTHUMP00000238557, S152, T cell antivation antigen S152, T-cell activation antigen CD27, T14, TNFRSF 7, TNFSF 7, Tp 55, Tumor necrosis factor receptor superfamily member 7
- WB
Lab
Western blot - Human CD27A knockout Raji cell line (AB274910)
ab133761 was shown to react with CD27 in wild-type Raji cells in western blot. Loss of signal was observed when CD27 knockout cell line ab274910 (knockout cell lysate ab281360) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab133761 overnight at 4 ° at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1 in 5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.
All lanes:
Western blot - Anti-CD27 antibody [EPR8568] (<a href='/en-us/products/primary-antibodies/cd27-antibody-epr8568-ab133761'>ab133761</a>) at 1/1000 dilution
Lane 1:
Wild-type Raji cell lysate at 20 µg
Lane 2:
CD27 knockout Raji cell lysate at 20 µg
Lane 2:
Western blot - Human CD27A knockout Raji cell line (ab274910)
Lane 3:
Ramos cell lysate at 20 µg
Lane 4:
Human Brain tissue lysate at 20 µg
Predicted band size: 29 kDa
Observed band size: 50 kDa
false
Exposure time: 2min
- WB
Lab
Western blot - Human CD27A knockout Raji cell line (AB274910)
Lanes 1 - 4 : Merged signal (red and green). Green - ab131254 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab131254 was shown to react with CD27 in wild-type Raji cells in western blot with loss of signal observed in CD27 knockout sample. Wild-type and CD27 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab131254 and 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 (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-CD27 antibody [EPR8569] (<a href='/en-us/products/primary-antibodies/cd27-antibody-epr8569-ab131254'>ab131254</a>) at 1/1000 dilution
Lane 1:
Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 2:
CD27 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human CD27A knockout Raji cell line (ab274910)
Lane 3:
Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 4:
Human brain tissue lysate at 20 µg
Predicted band size: 29 kDa
Observed band size: 35 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human CD27A knockout Raji cell line (AB274910)
1 bp mismatch after Lys57 and 10 bp deletion after Cys61 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human CD27A knockout Raji cell line (AB274910)
Knockout achieved by CRISPR/Cas9; X = 10 bp deletion; Frameshift : 100%
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
One finds CD27 at the heart of cellular immune responses. It does not function alone; instead it typically forms part of a receptor-ligand complex with CD70 which influences T cell and B cell activities. This interaction promotes cell survival and plays a role in the activation of NF-kB and MAPK8/JNK signaling pathways which are essential for immune response regulation. These pathways ensure that the body's immune cells can efficiently respond to pathogens.
Pathways
CD27 actively integrates into the NF-kB and mTOR signaling pathways which are important for regulating immune and inflammatory responses. Through these pathways CD27 works closely with proteins like TRAF2 and TRAF3 which are adapter molecules that transmit signals from the receptor. These signaling pathways provide a framework for understanding how CD27 and associated proteins work together to control immune cell behavior facilitating coordinated immune defense mechanisms.
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
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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