Mouse Fc gamma RIIIB/CD16b Antibody

Catalog #: MAB11725 Datasheet / COA / SDS

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MAB11725-100
MAB11725-SP

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Detection of CD16b in Mouse Colon viaseqIF™ staining on COMET™

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Mouse Fc gamma RIIIB/CD16b Antibody Summary

Species Reactivity

Mouse

Specificity

Detects a synthetic peptide specific for mouse CD16b around amino acid 60 in Direct ELISA.

Source

Monoclonal Rat IgG_2B Clone # 1107512

Purification

Protein A or G purified from hybridoma culture supernatant

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

Mouse liver tissue

Immunohistochemistry

3-25 µg/mL

Perfusion fixed paraffin-embedded sections of mouse liver and thymus

Multiplex Immunofluorescence

10 µg/mL

Perfusion fixed paraffin-embedded sections of Mouse Colon, Spleen, Thymus and Stomach

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

Multiplex Immunofluorescence

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Detection of CD16b in Mouse Colon viaseqIF™ staining on COMET™ CD16b was detected in perfusion fixed paraffin-embedded sections of mouse Colon using Rat Anti-Mouse CD16b, Monoclonal Antibody (Catalog #MAB11725) at 10ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module)and Dewax and HIER Buffer H (pH 9; Epredia Catalog #TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100).Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Multiplex Immunofluorescence

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Detection of CD16b in Mouse Spleen viaseqIF™ staining on COMET™ CD16b was detected in perfusion fixed paraffin-embedded sections of mouse Spleen using Rat Anti-Mouse CD16b, Monoclonal Antibody (Catalog #MAB11725) at 10ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module)and Dewax and HIER Buffer H (pH 9; Epredia Catalog #TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100).Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Multiplex Immunofluorescence

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CD16b Antibody in Mouse Thymus viaseqIF™ staining on COMET™ CD16b was detected in perfusion fixed paraffin-embedded sections of mouse Thymus using Rat Anti-Mouse CD16b, Monoclonal Antibody (Catalog #MAB11725) at 10ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module)and Dewax and HIER Buffer H (pH 9; Epredia Catalog #TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100).Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Multiplex Immunofluorescence

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Detection of CD16b in Mouse Stomach viaseqIF™ staining on COMET™ CD16b was detected in perfusion fixed paraffin-embedded sections of mouse Stomach using Rat Anti-Mouse CD16b, Monoclonal Antibody (Catalog #MAB11725) at 10ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module)and Dewax and HIER Buffer H (pH 9; Epredia Catalog #TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 555 Goat anti-Rat IgG Secondary Antibody at 1:100 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR555RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100).Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Western Blot

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Detection of Mouse Fc gamma RIIIB/CD16b by Western Blot. Western Blot shows lysates of mouse liver tissue. PVDF membrane was probed with 2 µg/ml of Rat Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11725) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Fc gamma RIIIB/CD16b at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Immunohistochemistry

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Detection of Fc gamma RIIIB/CD16b in Mouse Liver. Fc gamma RIIIB/CD16b was detected in perfusion fixed paraffin-embedded sections of mouse liver using Rat Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11725) at 5 µg/ml overnight at 4 °C. 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 the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Immunohistochemistry

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Detection of Fc gamma RIIIB/CD16b in Mouse Thymus. Fc gamma RIIIB/CD16b was detected in perfusion fixed paraffin-embedded sections of mouse thymus using Rat Anti-Mouse Fc gamma RIIIB/CD16b Monoclonal Antibody (Catalog # MAB11725) at 5 µg/ml overnight at 4 °C. 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 the HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

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).

References

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.

Long Name

Fc gamma Receptor III

Entrez Gene IDs

2215 (Human); 14131 (Mouse)

Alternate Names

CD16; CD16b antigen; CD16b; Fc fragment of IgG, low affinity IIIb, receptor (CD16b); Fc fragment of IgG, low affinity IIIb, receptor for (CD16); Fc gamma RIIIB; FCG3; Fc-gamma receptor IIIb (CD 16); Fc-gamma RIII; Fc-gamma RIIIb; Fc-gamma RIII-beta; FCGR3; FCGR3B; FcgRIIIB; FcR-10; fcRIII; FCRIIIB; IGFR3; IgG Fc receptor III-1; low affinity immunoglobulin gamma Fc region receptor III-B