Invitrogen

CD16/CD32 Monoclonal Antibody (93), PerCP-eFluor 710, eBioscience™

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CD16/CD32 Antibody — Fig. 5 Enforced expression of DAB2 predisposes mice to a serially transplantable myeloid malignancy. a Schematic of the primary transplant experiment analyzed in b through g . CBC complete blood counts, LDA limiting dilution assay. b A Kaplan-Meier survival curve was generated for mice receiving transplanted marrow containing Vector or DAB2 constructs following primary (1deg DAB2) or secondary transplant (2deg DAB2) in those mice dying of myeloproliferation or leukemia. c Peripheral blood analysis of white blood cells (WBCs), platelets (Plt), mean cell volume (MCV), and hemoglobin (HGB) at 25 weeks post transplant (mean +- SEM, Vector n = 9, 1deg DAB2 n = 16, 1deg DAB2-AML/MPD n = 7, 2deg DAB2 n = 8). d Spleen weights were assessed in primary and secondary recipients (mean +- SEM, Vector n = 6, 1deg DAB2-AML/MPD n = 6, 2deg DAB2-AML/MPD n = 8). e CFU analysis was performed using primary DAB2 -AML/MPD marrow sorted for GFP (mean +- SEM, GFP- BM n = 3, GFP + BM n = 4). f Frequency of LSK (Lin - Sca1 + c-Kit + ), CMP (Lin - Sca1 - c-Kit + CD34 + CD16/32 lo ), GMP (Lin - Sca1 - c-Kit + CD34 + CD16/32 hi ) and MEP (Lin - Sca1 - c-Kit + CD34 - CD16/32 lo ) determined by flow cytometry in GFP + marrow of 1deg DAB2 mice 60 weeks post transplant that did not develop leukemia or a myeloproliferative disorder (mean +- SEM, Vector n = 3-5, 1deg DAB2 n = 3-4). g Secondary limiting dilution assays using marrow from long-term engrafted Vector or DAB2 mice (52 weeks post-primary transplant).

CD16/CD32 Antibody in Flow Cytometry (Flow) — Staining of C57BL/6 splenocytes with Anti-Mouse CD45RB APC (Product # 17-0455-81) and 0.125 µg of Rat IgG2a K Isotype Control PerCP-eFluor® 710 (Product # 46-4321-82) (left) or 0.06 µg of Anti-Mouse CD16/CD32 PerCP-eFluor® 710 (right). Total viable cells were used for analysis.

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

0.125 µg/test

Published Applications

Flow Cytometry (Flow)

See 15 publications below

Product Specifications

Species Reactivity

Mouse

Published species

Fruit fly, Mouse

Host / Isotype

Rat / IgG2a, lambda

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), PerCP-eFluor 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

Conjugate

PerCP-eFluor™ 710

Excitation/Emission Profile

View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.1% gelatin

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

RRID

AB_2016701

Product Specific Information

Description: The 93 monoclonal antibody reacts with an epitope shared by mouse CD16 and CD32. CD16 (Fc gamma III Receptor) and CD32 (Fc gamma II Receptor) are the low affinity receptors for the mouse IgG Fc portion and are expressed by B cells, monocyte/macrophages, NK cells, and neutrophils.

Applications Reported: This 93 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 93 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 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: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

CD16 is a 50-65 kDa cell surface molecule that exists in two forms - a transmembranous form expressed by natural killer (NK) cells, and some T cells and a phosphatidylinositol linked form expressed by granulocytes. CD16 is a low affinity receptor for IgG (FcR III), and is an important receptor mediating ADCC (antibody dependent cell mediated cytotoxicity) by NK cells. IN comparison, CD32 is a 40 kD glycoprotein that acts as a low affinity receptor for IgG (also known as Fc gamma RII), and mediates several functions including endocytosis, activation of secretion, cytotoxicity and immunomodulation. CD32 is expressed by B cells, monocytes, granulocytes and platelets. Further, CD32 is involved in the phagocytosis of immune complexes, the regulation of antibody production by B-cells, and variations of the CD32 gene may increase vulnerability to systemic lupus erythematosus (SLE). Several transcript variants encoding different isoforms of CD32 have been found.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: CD16A; CD16B; CD32; CD32 receptor 2; Fc gamma Receptor 2; Fc gamma Receptor 3; Fc gamma Receptor II; Fc gamma receptor IIB; Fc gamma Receptor III; Fc gamma RIIB; Fc-gamma RII; Fc-gamma-RII; fc-gamma-RIIB; FCG2; FcgammaRIIB; FCGR2; FCGR2A; FCGR2B; FCGR2C; FCGR3; FCGR3A; FCGR3B; FCGRIII; FCR-10; FcRII-b; FcRII-c; FCRIII; FCRIIIA; FGFR2B; IgG Fc receptor II beta; Low affinity immunoglobulin gamma Fc region receptor II; low affinity immunoglobulin gamma Fc region receptor II-b; Ly-17; Lymphocyte antigen 17; RP11-5K23.1

Gene Aliases: AI528646; CD32; F630109E10Rik; Fc[g]RII; Fcgr2; Fcgr2a; Fcgr2b; FcgRII; Fcr-2; Fcr-3; fcRII; Ly-17; Ly-m20; LyM-1

UniProt ID: (Mouse) P08101

Entrez Gene ID: (Mouse) 14130

Function(s)

Process(es)

receptor signaling protein activity protein binding IgG binding immunoglobulin binding

CD16/CD32 Monoclonal Antibody (93), PerCP-eFluor 710, eBioscience™

Product Details

Product Specific Information

Target Information

Bioinformatics

Material safety data sheets (MSDS)

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