CD27 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 = 10 bp deletion Frameshift: 100%.
Images
Key facts
- Cell type
- Raji
- Species or organism
- Human
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 10 bp deletion Frameshift: 100%
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
Recommended products
CD27 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 = 10 bp deletion Frameshift: 100%.
Key facts
- Cell type
- Raji
- Species or organism
- Human
- Tissue
- Lymphatic
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9 X = 10 bp deletion Frameshift: 100%
- Disease
- Lymphoma
- Concentration
Properties
- Gene name
- CD27
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Suspension
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
CD27 also known as Tumor Necrosis Factor Receptor Superfamily Member 7 (TNFRSF7) is a protein chiefly involved in T cell activation and proliferation. It belongs to the TNF-receptor superfamily and weighs approximately 50 kDa. CD27 is expressed mainly on T lymphocytes and to a lesser extent on B cells and natural killer cells. Researchers often study CD27 and its ligand CD70 to understand immune responses. Tools such as anti-CD27 percp 7h21 monoclonal antibodies and APC-conjugated forms help detect this protein in various assays.
- 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.
- Associated diseases and disorders
CD27 has significant implications in autoimmune diseases and cancer. For example aberrant activation of CD27 may contribute to conditions like lymphoma and autoimmune diseases such as lupus. The CD70-CD27 signaling axis when dysregulated can lead to uncontrolled cell proliferation or apoptosis linking it to these pathologies. Understanding the interplay between CD27 and proteins like Bcl-2 which influences apoptosis helps pave the way for therapeutic strategies targeting these diseases.
Product promise
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4 product images
Western blot - Human CD27A knockout Raji cell line (ab274910)
Anti-CD27 antibody [EPR8568] 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 Human CD27A knockout Raji cell lysate ab281360) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with Anti-CD27 antibody [EPR8568] 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 (Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaging.
All lanes: Western blot - Anti-CD27 antibody [EPR8568] (Anti-CD27 antibody [EPR8568] ab133761) 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
Performed under reducing conditions.
Predicted band size: 29 kDa
Observed band size: 50 kDa
Exposure time: 2min
Western blot - Human CD27A knockout Raji cell line (ab274910)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD27 antibody [EPR8569] ab131254 observed at 35 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.Anti-CD27 antibody [EPR8569] 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 Anti-CD27 antibody [EPR8569] ab131254 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 hour at room temperature before imaging.
All lanes: Western blot - Anti-CD27 antibody [EPR8569] (Anti-CD27 antibody [EPR8569] ab131254) 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
Performed under reducing conditions.
Predicted band size: 29 kDa
Observed band size: 35 kDa
Next Generation Sequencing - Human CD27A knockout Raji cell line (ab274910)
Knockout achieved by CRISPR/Cas9; X = 10 bp deletion; Frameshift: 100%
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
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