FCGR1A KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 2 bp deletion; Frameshift: 99.99%.
Images
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 2 bp deletion; Frameshift: 99.99%
Alternative names
CD64 antigen, CD64A, CD64b, CD64c, FCG 1, FCGR1A, FCGR1B, FCGR1C, FCGR1_HUMAN, FCRIC, FLJ18345, Fc fragment of IgG high affinity Ia receptor, Fc fragment of IgG high affinity Ib receptor, Fc fragment of IgG high affinity Ic receptor, Fc fragment of IgG, high affinity Ia, receptor (CD64), Fc fragment of IgG, high affinity Ia, receptor for, Fc fragment of IgG, high affinity Ia, receptor for (CD64), Fc fragment of IgG, high affinity Ib, receptor (CD64), Fc fragment of IgG, high affinity Ib, receptor for, Fc fragment of IgG, high affinity Ic, receptor (CD64), Fc fragment of IgG, high affinity Ic, receptor (CD64), pseudogene, Fc fragment of IgG, high affinity Ic, receptor for, Fc gamma RIB, Fc gamma receptor, Fc gamma receptor I, Fc gamma receptor I A1, Fc gamma receptor I B2, Fc of IgG high affinity Ia receptor, Fc-gamma RI, Fc-gamma RIA, Fc-gamma RIC, FcRI, FcRIB, HFcgammaRIB, High affinity immunoglobulin gamma Fc receptor I, High affinity immunoglobulin gamma Fc receptor IB, IGFRB, IGFRC, IgG Fc receptor I, IgG Fc receptor IB, IgG Fc receptor IC, Immunoglobulin G Fc receptor I, Immunoglobulin G Fc receptor IB, Immunoglobulin G Fc receptor IC, MGC137713
Recommended products
FCGR1A KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 2 bp deletion; Frameshift: 99.99%.
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 2 bp deletion; Frameshift: 99.99%
- Disease
- Acute Monocytic Leukemia
- Concentration
Properties
- Gene name
- FCGR1A
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- 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 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.
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
CD64 also known as Fc gamma receptor 1a (FCGR1A) is a high-affinity receptor for the Fc region of immunoglobulin G (IgG). This receptor weighs approximately 72 kDa and is expressed on the surface of immune cells such as macrophages monocytes and activated neutrophils. CD64 protein plays an important role in immune responses by binding to antibodies and mediating processes such as phagocytosis and antibody-dependent cellular cytotoxicity (ADCC). Researchers often use anti-CD64 antibodies and CD64 markers to study its expression in various cellular contexts especially in flow cytometry.
- Biological function summary
CD64 expression involves the binding of IgG which serves as a pivotal mechanism to facilitate phagocytosis and clearance of opsonized pathogens. CD64 does not operate solely; it is part of a larger receptor family that includes other Fc gamma receptors like CD16 and CD32. The regulation of CD64 differs from its relatives due to its higher affinity for IgG. This attribute enables CD64 to play a more significant role in triggering immune responses especially during infections and inflammatory processes.
- Pathways
CD64 participates in the immune response pathway and is critical in the regulation of inflammatory responses. Key proteins interacting with CD64 in these pathways include the aforementioned CD16 and CD32 forming a network that ensures efficient immune system activation. The interaction with IgG and subsequent signal transduction underpin the receptor's functionality in these pathways harmonizing innate and adaptive immune responses.
- Associated diseases and disorders
CD64 expression correlates strongly with autoimmune disorders and inflammatory conditions. The receptor's role in orchestrating immune cell activation makes it relevant in diseases such as rheumatoid arthritis and sepsis. In rheumatoid arthritis CD64 interacts with immune complexes intensifying inflammation through enhancement of phagocytic activity. In sepsis increased CD64 marker expression on neutrophils can serve as a diagnostic indicator. Related proteins like CD16 and CD32 also participate in these disorders by modulating immune complex handling and cellular activation.
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4 product images
Western blot - Human FCGR1A knockout THP-1 cell line (ab275843)
False colour image of Western blot: Anti-CD64 antibody [EPR4624] staining at 1/10000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-CD64 antibody [EPR4624] ab134073 was shown to bind specifically to CD64. A band was observed at 45-50/55-80 kDa in wild-type THP-1 cell lysates with no signal observed at this size in FCGR1A knockout cell line ab275843 (knockout cell lysate Human FCGR1A knockout THP-1 cell lysate ab275817). To generate this image wild-type and FCGR1A knockout THP-1 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were 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) at 1/20000 dilution.
All lanes: Western blot - Anti-CD64 antibody [EPR4624] (Anti-CD64 antibody [EPR4624] ab134073) at 1/10000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: FCGR1A knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human FCGR1A knockout THP-1 cell line (ab275843)
Lane 3: U937 cell lysate at 20 µg
Lane 4: HEK-293 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 45 kDa, 45-50 kDa
Western blot - Human FCGR1A knockout THP-1 cell line (ab275843)
False colour image of Western blot: Anti-CD64 antibody [EPR4623] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-CD64 antibody [EPR4623] ab109449 was shown to bind specifically to CD64. A band was observed at 45-50/55-80 kDa in wild-type THP-1 cell lysates with no signal observed at this size in FCGR1A knockout cell line ab275843 (knockout cell lysate Human FCGR1A knockout THP-1 cell lysate ab275817). To generate this image wild-type and FCGR1A knockout THP-1 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were 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) at 1/20000 dilution.
All lanes: Western blot - Anti-CD64 antibody [EPR4623] (Anti-CD64 antibody [EPR4623] ab109449) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: FCGR1A knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human FCGR1A knockout THP-1 cell line (ab275843)
Lane 3: U937 cell lysate at 20 µg
Lane 4: HEK-293 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 43 kDa
Observed band size: 45-50 kDa
Next Generation Sequencing - Human FCGR1A knockout THP-1 cell line (ab275843)
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion, 2 bp deletion; Frameshift: 99.99%
Next Generation Sequencing - Human FCGR1A knockout THP-1 cell line (ab275843)
1 bp (allele 1) or 2 bp deletion (allele 2) after Asn 77 of the WT protein
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