We used the CyTOF for high-dimensional analysis of cell surface markers, cytokines, and signaling molecules simultaneously at the single-cell level. A panel of 37 metal-labeled antibodies (CD45, CD16/32, CD64, CD11b, CD11c, CD206, CD80, CD86, MHC-II (I-A/I-E), MHC-I, CX3CR1, F4/80, CD169, Ly6G, Ly6C, CD19, CD4, CD8, NK1.1, Epcam, TNFα, IL-10, and active Caspase-3 were purchased from FLUDIGM (Table S2 ). Primary antibodies for SiglecF, MERTK, Tim4, Marco, CD24, Arginase-1 (Arg1), CCR2, CD68, V-ATPase, Anti-NOX2, Lyve1, CD163, CD31, CD103, and BrdU were purchased from BD Bioscience, R&D Biosystems, BioLegend, and Thermo Fischer (Table S2 ), and labeled with FLUDIGM metal labeling kit (Maxpar X8 metal Labeling Kit) according to the manufacturer’s protocol. Cells were treated with Golgistop/GolgiPlug (BD Bioscience) for 2–3 h at 37°C and stained according to the FLUIDIGM recommended protocol. Samples were run on the Helios CyTOF mass cytometer (FLUIDIGM) at the flow cytometry core of the Research Resources Center of the University of Illinois at Chicago. Data from CyTOF were analyzed using the Cytobank online analysis tool (https://www.cytobank.org ).
Lyve1
Manufactured by BD
Sourced in United Kingdom
Lyve1 is a laboratory equipment product designed for use in scientific research. It serves as a tool for the detection and analysis of specific biomolecules. The core function of Lyve1 is to facilitate the identification and quantification of target analytes in samples.
Automatically generated - may contain errors
Lab products found in correlation
Helios cytof mass cytometer, by Standard BioTools (1 mentions)
Golgiplug golgistop, by BD (1 mentions)
Cd103, by BD (1 mentions)
Cd163, by BD (1 mentions)
Anti nox2, by BD (1 mentions)
Bromodeoxyuridine (brdu), by BD (1 mentions)
Siglec f, by BD (1 mentions)
Ds ri1 digital camera, by Nikon (1 mentions)
Eclipse te2000 u inverted microscope, by Nikon (1 mentions)
Hoechst 33342, by Merck Group (1 mentions)
2 protocols using lyve1
1
High-Dimensional CyTOF Analysis of Cell Markers
2
Fluorescent Staining and Imaging Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Fluorescein diacetate (FdA) live cell staining was performed according to the
protocol previously published.31 (link)
Histological hematoxylin/eosin and immunofluorescence staining on tissue
sections were described in detail by Biedermann et al.15 (link)
and whole mount staining by Marino et al.30
All the used antibodies were specific for human antigens if not
differently specified: from Abcam (Cambridge, UK): Lyve1 (polyclonal, 1:100,
ab10278); from BD Pharmingen (Basel, Switzerland): Ki67 (clone: B56, 1:100);
from Dako: human CD31 (clone: JC70A, 1:50), rat CD31 (clone: TLD-3A12, 1:50),
CK10 (clone: DE-K10, 1:100), Melanosome (HMB45, 1:50); from Dianova (Germany):
CD90 (clone: AS02, 1:50); from Lubio (Luzern, Switzerland): CK1 (clone: LHK1,
1:200); from Progen (Heidelberg, Germany): CK5 (polyclonal, 1:100, GP-CK5); from
SantaCruz (Labforce AG, Nunningen, Switzerland): Laminin 332 (clone: P3H9-2,
1:100), Tyrosinase (clone: c-19, 1:50), VE-Cadherin (clone: F-8, 1:50); from
Spring Bioscience (CA, Pleasanton): CK15 (clone: SPM190, 1:50). The
counterstaining of cell nuclei was performed with Hoechst 33342 (Sigma-Aldrich).
Pictures were taken with a DS-Ri1 digital camera connected to a Nikon Eclipse
TE2000-U inverted microscope. The device is equipped with Hoechst-, FITC-, and
TRITC-filter sets (Nikon AG, Switzerland; Software: NIS-Elements BR version
3.22.11).
protocol previously published.31 (link)
Histological hematoxylin/eosin and immunofluorescence staining on tissue
sections were described in detail by Biedermann et al.15 (link)
and whole mount staining by Marino et al.30
All the used antibodies were specific for human antigens if not
differently specified: from Abcam (Cambridge, UK): Lyve1 (polyclonal, 1:100,
ab10278); from BD Pharmingen (Basel, Switzerland): Ki67 (clone: B56, 1:100);
from Dako: human CD31 (clone: JC70A, 1:50), rat CD31 (clone: TLD-3A12, 1:50),
CK10 (clone: DE-K10, 1:100), Melanosome (HMB45, 1:50); from Dianova (Germany):
CD90 (clone: AS02, 1:50); from Lubio (Luzern, Switzerland): CK1 (clone: LHK1,
1:200); from Progen (Heidelberg, Germany): CK5 (polyclonal, 1:100, GP-CK5); from
SantaCruz (Labforce AG, Nunningen, Switzerland): Laminin 332 (clone: P3H9-2,
1:100), Tyrosinase (clone: c-19, 1:50), VE-Cadherin (clone: F-8, 1:50); from
Spring Bioscience (CA, Pleasanton): CK15 (clone: SPM190, 1:50). The
counterstaining of cell nuclei was performed with Hoechst 33342 (Sigma-Aldrich).
Pictures were taken with a DS-Ri1 digital camera connected to a Nikon Eclipse
TE2000-U inverted microscope. The device is equipped with Hoechst-, FITC-, and
TRITC-filter sets (Nikon AG, Switzerland; Software: NIS-Elements BR version
3.22.11).
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