Mouse Fc gamma RIIIB/CD16b Antibody
Mouse Fc gamma RIIIB/CD16b Antibody Summary
Ala31-Thr215
Accession # P08508
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of CD16 in Mouse Spleen via seqIF™ staining on COMET™ CD16 was detected in perfusion fixed frozen sections of mouse Spleen using Rabbit Anti-Mouse CD16, Monoclonal Antibody (Catalog #MAB11667) at 15ug/mL at 27 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5 minutes at room temperature followed by a 20-minute incubation inTris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm and membrane. Protocol available in COMET™ Panel Builder.
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Detection of Mouse Fc gamma RIIIB/CD16b by Western Blot. Western Blot shows lysates of mouse spleen tissue. PVDF membrane was probed with 2 µg/ml of Rabbit Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11667) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Fc gamma RIIIB/CD16b at approximately 40-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
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Detection of Fc gamma RIIIB/CD16b in Mouse Liver. Fc gamma RIIIB/CD16b was detected in immersion fixed paraffin-embedded sections of mouse liver using Rabbit Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11667) at 0.1 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cell surface of Kupffer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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Detection of Fc gamma RIIIB/CD16b in Mouse Lung. Fc gamma RIIIB/CD16b was detected in immersion fixed paraffin-embedded sections of mouse lung using Rabbit Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11667) at 0.5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cell surface of macrophages. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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Detection of Fc gamma RIIIB/CD16b in Mouse Thymus. Fc gamma RIIIB/CD16b was detected in immersion fixed paraffin-embedded sections of mouse thymus using Rabbit Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11667) at 0.5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003) or the HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cell surface. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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Detection of Fc gamma RIIIB/CD16b in RAW264.7 cells Fc gamma RIIIB/CD16b was detected in immersion fixed RAW264.7 mouse monocyte/macrophage cell line (Positive) and absent in NIH‑3T3 mouse embryonic fibroblast cell line (Negative) using Rabbit Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11667) at 3 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to the cell surface of RAW264.7 (positive) cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Fc gamma RIIIB/CD16b
Receptors for the Fc region of IgG (Fc gamma Rs) are members of the Ig superfamily that function in the activation or inhibition of immune responses such as degranulation, phagocytosis, ADCC (antibody‑dependent cellular toxicity), cytokine release, and B cell proliferation (1-3). The Fc gamma Rs have been divided into three classes based on close relationships in their extracellular domains; these groups are designated Fc gamma RI (also known as CD64), Fc gamma RII (CD32), and Fc gamma RIII (CD16). Each group may be encoded by multiple genes and exist in different isoforms depending on species and cell type. The CD64 proteins are high affinity receptors (~10-8-10-9 M) capable of binding monomeric IgG, whereas the CD16 and CD32 proteins bind IgG with lower affinities (~10-6-10-7 M) only recognizing IgG aggregates surrounding multivalent antigens (1, 4). Fc gamma Rs that deliver an activating signal either have an intrinsic immunoreceptor tyrosine-based activation motif (ITAM) within their cytoplasmic domains or associate with one of the ITAM-bearing adapter subunits, Fc R gamma or zeta (3, 5). The only inhibitory member in human and mouse, Fc gamma RIIb, has an intrinsic cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM). The coordinated functioning of activating and inhibitory receptors is necessary for successful initiation, amplification, and termination of immune responses (5).
Mouse CD16 is encoded by a single gene. The protein product is a type I transmembrane protein having two extracellular Ig-like domains. It is expressed on a variety of myeloid and lymphoid cells (4) and associates with Fc R gamma to deliver an activating signal upon ligand binding (5). Mouse CD32 is closely related to mouse CD16 throughout its extracellular domain (95% amino acid sequence identity), but has a divergent cytoplasmic domain and functions as an inhibitory receptor. Together these proteins constitute an activating/inhibiting receptor pair to regulate immune responses (5).
- van de Winkel, J. and P. Capes (1993) Immunol. Today 14:215.
- Raghaven, M. and P. Bjorkman (1996) Annu. Rev. Cell Dev. Biol. 12:181.
- Ravetch, J. and S. Bolland (2001) Annu. Rev. Immunol. 19:275.
- Takai, T. (2002) Nature Rev. Immunol. 2:580.
- Ravetch, J. and L. Lanier (2000) Science 290:84.
Product Datasheets
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