Human CCR2 knockout THP-1 cell line
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- Flow Cyt
Supplier Data
Flow Cytometry - Human CCR2 knockout THP-1 cell line (AB273716)
Flow cytometry overlay histogram showing wild-type THP-1 (green line) and CCR2 knockout THP-1 cells (ab273716, red line) stained with ab227236. The cells were incubated in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab227236) (1x106 in 100μl at 1 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 THP-1 - black line; CCR2 THP-1 knockout - 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.
- Sanger seq
Supplier Data
Sanger Sequencing - Human CCR2 knockout THP-1 cell line (AB273716)
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Cell Lines & Lysates
AB273760
Human CCL3 (Macrophage Inflammatory Protein 1) knockout THP-1 cell line
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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 bath for 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 cells/mL. 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.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
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
CCR2 plays an important role in mediating leukocyte migration. It acts in the immune system to guide monocytes from the bloodstream into tissues contributing to immune surveillance and response. CCR2 operates not as part of a larger receptor complex but it does interact closely with other chemokine receptors which may influence its signaling. The receptor's activity has critical implications for inflammatory processes and lies at the heart of many immune responses.
Pathways
CCR2 is integrally involved in the chemokine signaling pathway and the inflammatory response pathway. Its function in these pathways highlights its role in modulating immune cell infiltration during immune challenges. The receptor also interfaces with other important signaling proteins such as CCR5 which like CCR2 is another chemokine receptor involved in mediating immune cell movement. These interactions overlap and complement each other offering nuanced regulation of immune cell dynamics.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
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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