Human CXCR5 knockout Raji cell line
- Advanced Validation
- What is this?
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(0 Publication)
- Flow Cyt
Lab
Flow Cytometry - Human CXCR5 knockout Raji cell line (AB273380)
Flow cytometry overlay histogram showing wild-type Raji (green line) and CXCR5 knockout Raji cells (ab273380) stained with ab254415 (red line). The cells were incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serumto block FC receptors and non-specific protein-protein interaction followed by the antibody (ab254415) (1x106 in 100μl at 0.2 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type Raji - black line; CXCR5 knockout Raji - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
- WB
Lab
Western blot - Human CXCR5 knockout Raji cell line (AB273380)
Lanes 1 - 4 : Merged signal (red and green). Green - ab254415 observed at 60 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab254415 was shown to react with CXCR5 in Raji wild-type cells in Western blot with loss of signal observed in CXCR5 knockout sample. Wild-type and CXCR5 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab254415 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 h at room temperature before imaging.
All lanes:
Western blot - Anti-CXCR5 antibody [EPR23463-30] (<a href='/en-us/products/primary-antibodies/cxcr5-antibody-epr23463-30-ab254415'>ab254415</a>) at 1/1000 dilution
Lane 1:
Wild-type Raji cell lysate at 30 µg
Lane 2:
CXCR5 knockout Raji cell lysate at 30 µg
Lane 2:
Western blot - Human CXCR5 knockout Raji cell line (ab273380)
Lane 3:
Daudi cell lysate at 30 µg
Lane 4:
Jurkat cell lysate at 30 µg
Predicted band size: 42 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human CXCR5 knockout Raji cell line (AB273380)
Homozygous : 19 bp deletion in exon 2
Complementary Products
Primary Antibodies
AB254415
Anti-CXCR5 antibody [EPR23463-30]
BOND RX™ Validated|RabMAb|Recombinant|KO Validated
![Flow Cytometry - Anti-CXCR5 antibody [EPR23463-30] (AB254415)](https://content.abcam.com/products/images/cxcr5-antibody-epr23463-30-ab254415--flow-cytometry-img13375.jpg)
- 3
- 2
Primary Antibodies
AB64088
BOND RX™ Validated|20ul selling size|RabMAb|Advanced Validation|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [SP32] (AB64088)](https://content.abcam.com/products/images/cd20-antibody-sp32-ab64088--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img449343.jpg)
- 5
- 12
Primary Antibodies
AB246518
Anti-CXCL13 antibody [EPR23400-92]
BOND RX™ Validated|RabMAb|Recombinant
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL13 antibody [EPR23400-92] (AB246518)](https://content.abcam.com/products/images/cxcl13-antibody-epr23400-92-ab246518--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img81627.jpg)
- 5
- 1
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
Zygosity
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
CXCR5 serves a significant role in directing B cells and Tfh cells to lymphoid tissues by interacting with its ligand CXCL13. This interaction involves recruitment and organization of B cells into follicles within secondary lymphoid organs. The presence of CXCR5 allows B cells and Tfh cells to navigate the follicular zones efficiently enhance antigen-specific immune responses and contribute to long-lived humoral immunity. CXCR5 functions as part of a complex that includes signaling pathways influencing cellular movements and location.
Pathways
CXCR5 plays a significant part in the immune system's trafficking pathways particularly within the crosstalk between chemokines and their receptors. CXCR5 expression facilitates the CXC chemokine pathway which important for positioning immune cells within lymphoid organs. Additionally CXCR5 interacts with proteins such as CXCL13 working together to pattern the architecture of germinal centers where affinity maturation and class switching of antibodies occur critical processes in adaptive immunity.
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'.
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