47-0649-42

antibody from Invitrogen Antibodies

Targeting: FCGR1A CD64, CD64A

Provider product page for 47-0649-42

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [4]
Comments [0]
Validations
Flow cytometry [1]
Other assay [5]

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Product number: 47-0649-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD64 Monoclonal Antibody (10.1), APC-eFluor™ 780, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The 10.1 monoclonal antibody reacts with human CD64 (FcRI), a 75 kDa type I transmembrane protein. CD64 is the high affinity receptor for IgG and is involved in antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, and regulation of cytokine production. Monocytes and macrophages express CD64, while mature granulocytes and lymphocytes are negative. Applications Reported: This 10.1 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 10.1 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. APC-eFluor® 780 emits at 780 nm and is excited with the Red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 633-647 nm; Emission: 780 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 10.1
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

FCGR1A protein structure - 47-0649-42 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Murlentamab opsonization of SKOV3-R2 + orients naive macrophages and reprograms TAMs towards an M1-like profile. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). ( A ) The proportion of macrophages expressing M1/M2 membrane markers (CD32, CD64, CD80, TLR2, CD163, CD36 and CD206) was determined by flow cytometry after three days of co-culture with SKOV3-R2 + cells. ( B ) The release of cytokines (IL1beta, IL12, TNFalpha, IL6, IFNgamma, IL10) and chemokines (CCL2, CCL4, CCL5, CXCL9 and CXCL10) in the culture medium was determined by AlphaLISA after three days of co-culture with SKOV3-R2 + cells. Data shown (boxplots) are the results from three different experiments (performed with three different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey''s multiple comparisons test.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 CD64 and CD163 expression on monocytes, granulocytes, and lymphocytes.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Patient mutations lead to IFN-gamma-dependent defects in the iPSC-derived macrophages. ( A ) Representative flow cytometric analysis of surface marker (HLA-DR, CD64, CD38, CD282) upregulation after IFN-gamma stimulation in healthy and patient iPSC-derived macrophages. Blue: Isotype. Pink: Surface marker. ( B ) Fold change of median fluorescent intensity (MFI) of HLA-DR, CD64, CD38 and CD282 in healthy and patient iPSC-derived macrophages ( n = 3-7, mean +- SD; each dot represents macrophages from an independent harvest and from at least three independent differentiations, Kruskal-Wallis with Dunn''s multiple comparison)) ( C ) Representative western blot analysis of STAT1 phosphorylation (pSTAT1) after stimulation with low (25 ng/mL) or high (100 ng/mL) dose of IFN-gamma. Tubulin was used as a loading control. The left side of the blot shows the protein size marker. ( D ) Densitometric analysis of STAT1 phosphorylation after IFN-gamma stimulation. Values have been normalized to loading control. ( n = 3, mean +- SD; each dot represents macrophages from an independent harvest and from at least two independent differentiations, Kruskal-Wallis with Dunn''s multiple comparison). ( E ) qRT-PCR analysis of upregulation of downstream targets IRF1, CXCL10, CCL2 and CCL4 after IFNgamma stimulation. Values have been normalized to GAPDH as housekeeping gene. ( n = 3-5, mean +- SD; each dot represents macrophages from an independent harvest and from at least two independent diffe