Afs98

Manufactured by BioLegend

The AFS98 is a laboratory instrument designed for cell analysis and sorting. It is capable of detecting and quantifying multiple fluorescent markers simultaneously. The core function of the AFS98 is to enable researchers to accurately analyze and sort cell populations based on their specific characteristics.

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Lab products found in correlation

Facsaria, by BD (2 mentions) M1 70, by Thermo Fisher Scientific (2 mentions) M5 114, by Thermo Fisher Scientific (2 mentions) Ds5mmer, by Thermo Fisher Scientific (2 mentions) Rb6 8c5, by BioLegend (2 mentions) M1 70, by BioLegend (2 mentions) Lsrfortessa, by BD (1 mentions) Summit 4, by Agilent Technologies (1 mentions) Rb6 8c5, by Thermo Fisher Scientific (1 mentions) Rtk4530, by BioLegend (1 mentions) Facsdiva software, by BD (1 mentions) Fortessa, by BD (1 mentions) Af6 120, by BioLegend (1 mentions) Sa011f11, by BioLegend (1 mentions) Liberase, by Roche (1 mentions)

Close spelling variants of this product

CD115 (AFS98, (6 citations) CD115 (clone AFS98), (3 citations) BV-421–conjugated anti–mouse CD115 (clone AFS98, (2 citations) Anti-CD115 (AFS98; (2 citations) Anti-CD115-biotin (clone AFS98 (2 citations) Clone AFS98 (2 citations)

5 protocols using afs98

Sorting GM-DCs and GM-Macs from Mouse Bone Marrow

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BMDCs differentiated from the bone marrow of five C57BL/6J mice were
pooled and sorted on a BD FACS Aria (BD Biosciences). Briefly, BMDCs were
harvested, pooled, incubated with Fc receptor-blocking antibody (16-0161-86,
Thermo Fisher) for 15 minutes on ice, and the CD11c⁺ BMDCs were
enriched using CD11c microbeads following the manufacturer’s
instructions. To sort GM-DCs and GM-Macs, the CD11c-enriched BMDCs were
stained with antibodies against CD11c (N418, Thermo Fisher, 1:300), CD11b
(M1/70, Thermo Fisher, 1:300) MHC-II (M5/114.15.2, Thermo Fisher, 1:400),
CD115 (AFS98, Biolegend, 1:100), MerTK (DS5MMER, Thermo Fisher, 1:100), and
CD135 (A2F10, Biolegend, 1:50). GM-Macs were sorted as
CD11c⁺CD11b^hiMHCII^lo-intCD115⁺MerTK⁺while GM-DCs were sorted as
CD11c⁺CD11b^intMHCII^hiCD115⁻CD135⁺(see also Figure
S1). Dead cells were excluded with PI.

Donado C.A., Cao A.B., Simmons D.P., Croker B.A., Brennan P.J, & Brenner M.B. (2020). A Two-Cell Model for IL-1β Release Mediated by Death-Receptor Signaling. Cell reports, 31(1), 107466.

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Sorting GM-DCs and GM-Macs from Mouse Bone Marrow

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Donado C.A., Cao A.B., Simmons D.P., Croker B.A., Brennan P.J, & Brenner M.B. (2020). A Two-Cell Model for IL-1β Release Mediated by Death-Receptor Signaling. Cell reports, 31(1), 107466.

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Multicolor Flow Cytometry of Immune Cells

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100 µL of anti-coagulated (EDTA) whole blood was incubated with the appropriate antibody or isotype control in the dark at room temperature for 30 min: rat F4/80 Biolegend BM8 1∶200, rat Ly-6G/Ly-6C (Gr-1) Biolegend RB6-8C5 1∶200, rat CD115 (CSF1R) eBioscience AFS98 1∶1000, or Rat IgG2b, k Isotype Ctrl Biolegend RTK4530 1∶200). Blood was processed using DAKO Uti-lyse, erythrocyte lysing reagent (S3325) according to manufacturer's protocol. Samples were analyzed on BD LSR Fortessa (blood) and analysed using Summit 4.1 software (DAKO).

Sauter K.A., Pridans C., Sehgal A., Bain C.C., Scott C., Moffat L., Rojo R., Stutchfield B.M., Davies C.L., Donaldson D.S., Renault K., McColl B.W., Mowat A.M., Serrels A., Frame M.C., Mabbott N.A, & Hume D.A. (2014). The MacBlue Binary Transgene (csf1r-gal4VP16/UAS-ECFP) Provides a Novel Marker for Visualisation of Subsets of Monocytes, Macrophages and Dendritic Cells and Responsiveness to CSF1 Administration. PLoS ONE, 9(8), e105429.

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Multi-parameter Flow Cytometry Analysis

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Cells were stained with primary antibodies against murine CD45 (30-F11, Biolegend cat. no. 103112, RRID: AB_312976), CD11b (M1/70, Biolegend cat. no. 101205, RRID: AB_312788), F4/80 (CI:A3-1, Bio-Rad cat. no. MCA497GA), Ly6C/G (Gr-1) (RB6-8C5, Biolegend catno. 108411, RRID: AB_313376), Lyve1 (ALY7, Thermo Fisher Scientific cat. no. 53-0443-82, RRID: AB_1633415), I-Ab (AF6-120.1, Biolegend cat. no. 116421, RRID: AB_10613291), CD11c (N418, Biolegend cat. no. 117306, RRID: AB_313775), CD64 (X54-5/701, Biolegend cat. no. 139306, RRID: 11219391), CX 3 CR1 (SA011F11, Biolegend cat. no. 149015, RRID: AB_2565699), CD115 (AFS98, Biolegend cat. no. 135509, RRID: AB_2085222), Ly6C (HK1.4, Biolegend cat. no. 128015, RRID: AB_1732087). Methods adhered to published guidelines (Cossarizza et al., 2019) . Analysis was performed on a Fortessa (BD Biosciences), and data were acquired with FACSDiva software (BD Biosciences). Post-acquisition analysis was performed using FlowJo software (Tree Star, FlowJo LLC; Ashland, Oregon) .

Kim J.S., Kolesnikov M., Peled-Hajaj S., Scheyltjens I., Xia Y., Trzebanski S., Haimon Z., Shemer A., Lubart A., Van Hove H., Chappell-Maor L., Boura-Halfon S., Movahedi K., Blinder P, & Jung S. (2021). A Binary Cre Transgenic Approach Dissects Microglia and CNS Border-Associated Macrophages. Immunity, 54(1).

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Discriminating Tissue-Resident and Circulating Leukocytes

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Tissue-resident and circulation-borne leukocytes were discriminated by intravenous administration of an anti-CD45 antibody (30-F11, APC-Cy7, BioLegend). Two hours later mice were sacrificed. Blood was lysed with 3 ml lysis buffer/100 ml blood (lysis buffer: 150 mM NH 4 Cl; 10 mM KHCO 3 ; 0.1 mM diNaEDTA, pH 7.4). Aortas were digested with 1.25 mg/ml liberase (Roche) for 1 hour at 37 C. Cell suspensions were further processed for flow cytometry analysis. Blood leukocytes were stained with antibodies to CD45 (I3/2.3, BioLegend), CD11b (M1/70, BioLegend), CD115 (AFS98, BioLegend), Gr1 (RB6-8C5, BioLegend) and Ly6G (1A8, BioLegend) whereas aortic leukocytes were discriminated by antibodies to CD45 (I3/2.3, BioLegend), CD11b (M1/70, BioLegend), Gr1 (RB6-8C5, BioLegend) and Ly6G (1A8, BioLegend). Hearts were fixed in 4% paraformaldehyde for 24 hours before being incubated in 30% sucrose for 24 hours at 4 C. Afterwards hearts were embedded in Tissue-Tek (O.C.T., Sakura) and snap frozen. CD45 prelabelled leukocytes were visualized with an anti-rat IgG Dylight550 antibody (Thermo Fisher). To test vascular leakage we intravascular injected an anti-collagen type IV alpha 1 antibody (Novus Biologicals).

Winter C., Silvestre-Roig C., Ortega-Gomez A., Lemnitzer P., Poelman H., Schumski A., Winter J., Drechsler M., de Jong R., Immler R., Sperandio M., Hristov M., Zeller T., Nicolaes G.A.F., Weber C., Viola J.R., Hidalgo A., Scheiermann C, & Soehnlein O. (2018). Chrono-pharmacological Targeting of the CCL2-CCR2 Axis Ameliorates Atherosclerosis. Cell metabolism, 28(1).

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