Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [1]
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- Product number
- 25-0329-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD32 Monoclonal Antibody (6C4 (CD32)), PE-Cyanine7, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: This 6C4 monoclonal antibody reacts with human CD32 (also known as Fc gammaRII). This antibody recognizes two isoforms of the receptor, Fc gammaRIIA and Fc gammaRIIB. These 40-kDa polymorphic transmembrane glycoproteins are expressed on B cells, granulocytes, monocytes, macrophages, and platelets. Moreover, these receptors are detected on natural killer cells. CD32 enables interaction between Fc gammaRII-expressing cells and opsonized antigen or IgG-containing immune complexes. Both receptors exhibit low affinity towards IgG and play a role in inflammation and autoimmune disease. This clone has been reported to inhibit Ig binding in a rosette blocking assay. Applications Reported: This 6C4 (CD32) antibody has been reported for use in flow cytometric analysis. Applications Tested: This 6C4 (CD32) antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 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. 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 # 00-822-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-54) 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: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 6C4 (CD32)
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Staining of normal human peripheral blood cells with Anti-Human CD19 eFluor® 450 (Product # 48-0199-42) and Mouse IgG1 K Isotype Control PE-Cyanine7 (Product # 25-4714-80) (left) or Anti-Human CD32 PE-Cyanine7 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 6 Surface expression of key markers of microglia. a Flow cytometry analysis was used to measure the surface expression of CD11b, CD14, CD68, CD16, CD32, CD64, CD163, P2RY12, and TGFbetaR on the representative immortalized cell line hmuglia 1A1. In each experiment, 100,000 cells were resuspended in 1 mL of cold PBS in the presence of 0.5 mug of the antibody or isotype control for 20 min on ice. Appropriate secondary antibodies were used in the absence of fluorophore-conjugated primary antibody, and the cell-antibody complexes were centrifuged and resuspended in PBS. In each FACS profile, the gray distributions represent the proportion of cells bound to the isotype control, whereas the red distributions represent the proportions of cells bound by the target antibody. b Quantification of the abovementioned markers as well as GFAP, CD4, and CCR5 on the surface of primary human astrocytes and immortalized microglia, as indicated