Log In Sign up
The largest database of trusted experimental protocols

Maxpar x8 labeling kit

Manufactured by Standard BioTools
Visit Supplier
Sourced in United States

The MaxPar X8 labeling kit is a laboratory product designed for sample preparation and data acquisition in mass cytometry (CyTOF) workflows. The kit enables the conjugation of antibodies with metal-tagged polymers, allowing for the simultaneous measurement of up to 8 protein markers in a single sample.

Automatically generated - may contain errors

Lab products found in correlation

Iridium intercalator, by Standard BioTools (6 mentions) Cisplatin, by Standard BioTools (5 mentions) Cytof2, by Standard BioTools (5 mentions) Helios mass cytometer, by Standard BioTools (4 mentions) Foxp3 transcription factor fix perm buffer, by Thermo Fisher Scientific (2 mentions) Cisplatin, by Enzo Life Sciences (2 mentions) Facs lyse, by BD (2 mentions) Facs perm 2, by BD (2 mentions) 4 element normalization beads, by Standard BioTools (1 mentions) Cisplatin, by Merck Group (1 mentions) Permeabilization buffer, by Thermo Fisher Scientific (1 mentions) Eq beads, by Standard BioTools (1 mentions) Cd14 clone 3c10, by Thermo Fisher Scientific (1 mentions) Tcr vα7.2 clone 3c10, by BioLegend (1 mentions) Lsr fortessa 2, by BD (1 mentions) Ebioscience fixation permeabilization kit, by Thermo Fisher Scientific (1 mentions) Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions) Live dead fixable near ir, by Thermo Fisher Scientific (1 mentions) Trustain fcx, by BioLegend (1 mentions) Iridium interchelator, by Standard BioTools (1 mentions)

17 protocols using maxpar x8 labeling kit

1

Mass Cytometry Antibody Conjugation

Check if the same lab product or an alternative is used in the 5 most similar protocols
All anti-human antibodies pre-conjugated to metal isotopes were obtained from Fluidigm Corporation (San Francisco, US). All remaining antibodies were obtained from the indicated companies as purified antibodies and in-house conjugation was done using the MaxPar X8 labeling kit (Fluidigm). Table S2 shows a detailed list of all antibodies used for panel 1 and panel 2.
Schulte-Schrepping J., Reusch N., Paclik D., Baßler K., Schlickeiser S., Zhang B., Krämer B., Krammer T., Brumhard S., Bonaguro L., De Domenico E., Wendisch D., Grasshoff M., Kapellos T.S., Beckstette M., Pecht T., Saglam A., Dietrich O., Mei H.E., Schulz A.R., Conrad C., Kunkel D., Vafadarnejad E., Xu C.J., Horne A., Herbert M., Drews A., Thibeault C., Pfeiffer M., Hippenstiel S., Hocke A., Müller-Redetzky H., Heim K.M., Machleidt F., Uhrig A., Bosquillon de Jarcy L., Jürgens L., Stegemann M., Glösenkamp C.R., Volk H.D., Goffinet C., Landthaler M., Wyler E., Georg P., Schneider M., Dang-Heine C., Neuwinger N., Kappert K., Tauber R., Corman V., Raabe J., Kaiser K.M., Vinh M.T., Rieke G., Meisel C., Ulas T., Becker M., Geffers R., Witzenrath M., Drosten C., Suttorp N., von Kalle C., Kurth F., Händler K., Schultze J.L., Aschenbrenner A.C., Li Y., Nattermann J., Sawitzki B., Saliba A.E, & Sander L.E. (2020). Severe COVID-19 Is Marked by a Dysregulated Myeloid Cell Compartment. Cell, 182(6), 1419-1440.e23.
+ Open protocol
+ Expand
2

Metal-Labeling and CyTOF Analysis of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Metal-labeled Abs were obtained from Fluidigm or labeled using the MaxPar X8 labeling kit (Fluidigm) according to manufacturer’s instructions (see Table S1). Freshly isolated mouse (BSA- or Dynabead-enriched) splenocytes or thawed human mononuclear cells from blood and spleen were stained with 1 mL of 0.25 μM cisplatin (Fluidigm) for 5 minutes at room temperature to exclude dead cells. Cells were then washed with CyFACS buffer (2 mM EDTA, 1% BSA, 1% in PBS) and stained with heavy-metal-labeled Ab cocktail for 30 minutes on ice. Cells were washed twice with CyFACS then fixed with FoxP3 Transcription Factor Fix/Perm Buffer (Thermo Fisher Scientific) for 2 hours. Human surface CyTOF Abs that were sensitive to FoxP3 buffer in our hands (i.e., CX3CR1, CD123, CD33, CD135, CD172a and CD163), were instead stained after fixation and permeabilization for 30 minutes at 4°C. After staining, samples were washed and incubated with 2% paraformaldehyde (Electron) in PBS containing 125 nM Iridium intercalator (Fluidigm) overnight. Cells were washed with water, filtered, and acquired in a CyTOF2 (Fluidigm) at the Stanford Shared FACS Facility.
Leylek R., Alcántara-Hernández M., Lanzar Z., Lüdtke A., Perez O.A., Reizis B, & Idoyaga J. (2019). Integrated Cross-Species Analysis Identifies a Conserved Transitional Dendritic Cell Population. Cell reports, 29(11), 3736-3750.e8.
+ Open protocol
+ Expand
3

Cisplatin Staining and Mass Cytometry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were detached, collected, and stained with 5 µm Cisplatin (Sigma–Aldrich) for 5 min in a 37 °C water bath, and fixed using 1.6% Paraformaldehyde for 10 min at room temperature. Cells were then incubated for 10 min with blocking solution (BioLegend) and stained with a mixed antibody cocktail at room temperature for 30 min, followed by twice PBS washing. The antibodies are listed in Table S4 (Supporting Information). Metal‐labeled antibodies were labeled in‐house using the MaxPar X8 labeling kit according to the manufacturer's instructions (Fluidigm). Mass cytometry data were obtained using Helios (Fluidigm) as previously reported. Briefly, stained cells were suspended in deionized H2O, and added 4‐element normalization beads (Fluidigm) immediately before applying the samples to the instrument. Cells were acquired at a rate of 300–500 events s−1.
Xie H., Guo W., Jiang H., Zhang T., Zhao L., Hu J., Gao S., Song S., Xu J., Xu L., Sun X., Ding Y., Jiang L, & Ding X. (2024). Photosensitive Hydrogel with Temperature‐Controlled Reversible Nano‐Apertures for Single‐Cell Protein Analysis. Advanced Science, 11(19), 2308569.
+ Open protocol
+ Expand
4

Comprehensive Antibody Conjugation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
All anti-human antibodies pre-conjugated to metal isotopes were obtained from Fluidigm Corporation (San Francisco, USA). All remaining antibodies were obtained from the indicated companies as purified antibodies and in-house conjugation was done using the MaxPar X8 labeling kit (Fluidigm, USA). Antibodies are listed in the key resources table.
Georg P., Astaburuaga-García R., Bonaguro L., Brumhard S., Michalick L., Lippert L.J., Kostevc T., Gäbel C., Schneider M., Streitz M., Demichev V., Gemünd I., Barone M., Tober-Lau P., Helbig E.T., Hillus D., Petrov L., Stein J., Dey H.P., Paclik D., Iwert C., Mülleder M., Aulakh S.K., Djudjaj S., Bülow R.D., Mei H.E., Schulz A.R., Thiel A., Hippenstiel S., Saliba A.E., Eils R., Lehmann I., Mall M.A., Stricker S., Röhmel J., Corman V.M., Beule D., Wyler E., Landthaler M., Obermayer B., von Stillfried S., Boor P., Demir M., Wesselmann H., Suttorp N., Uhrig A., Müller-Redetzky H., Nattermann J., Kuebler W.M., Meisel C., Ralser M., Schultze J.L., Aschenbrenner A.C., Thibeault C., Kurth F., Sander L.E., Blüthgen N, & Sawitzki B. (2021). Complement activation induces excessive T cell cytotoxicity in severe COVID-19. Cell, 185(3), 493-512.e25.
+ Open protocol
+ Expand
5

Labeling Monoclonal Antibodies with Gadolinium

Check if the same lab product or an alternative is used in the 5 most similar protocols
Monoclonal antihuman HLA-DR (LN3, catalog no. 327002; BioLegend, San Diego, Calif) and fluorescein isothiocyanate-conjugated antihuman HLA-DR antibody (LN3, catalog no. ab1182; Abcam, Cambridge, England) were labeled with 160Gd by using the commercially available MAXPARX8 labeling kit (Fluidigm, San Francisco, Calif) according to the manufacturer’s instructions (manual PRD002v11) (18 (link)) (Appendix E1 [online]).
Savic L.J., Doemel L.A., Schobert I.T., Montgomery R.R., Joshi N., Walsh J.J., Santana J., Pekurovsky V., Zhang X., Lin M., Adam L., Boustani A., Duncan J., Leng L., Bucala R.J., Goldberg S.N., Hyder F., Coman D, & Chapiro J. (2020). Molecular MRI of the Immuno-Metabolic Interplay in a Rabbit Liver Tumor Model: A Biomarker for Resistance Mechanisms in Tumor-targeted Therapy?. Radiology, 296(3), 575-583.
+ Open protocol
+ Expand
6

Metal-Labeling and CyTOF Analysis of Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Metal-labeled Abs were obtained from Fluidigm or labeled using the MaxPar X8 labeling kit (Fluidigm) according to manufacturer’s instructions (see Table S1). Freshly isolated mouse (BSA- or Dynabead-enriched) splenocytes or thawed human mononuclear cells from blood and spleen were stained with 1 mL of 0.25 μM cisplatin (Fluidigm) for 5 minutes at room temperature to exclude dead cells. Cells were then washed with CyFACS buffer (2 mM EDTA, 1% BSA, 1% in PBS) and stained with heavy-metal-labeled Ab cocktail for 30 minutes on ice. Cells were washed twice with CyFACS then fixed with FoxP3 Transcription Factor Fix/Perm Buffer (Thermo Fisher Scientific) for 2 hours. Human surface CyTOF Abs that were sensitive to FoxP3 buffer in our hands (i.e., CX3CR1, CD123, CD33, CD135, CD172a and CD163), were instead stained after fixation and permeabilization for 30 minutes at 4°C. After staining, samples were washed and incubated with 2% paraformaldehyde (Electron) in PBS containing 125 nM Iridium intercalator (Fluidigm) overnight. Cells were washed with water, filtered, and acquired in a CyTOF2 (Fluidigm) at the Stanford Shared FACS Facility.
Leylek R., Alcántara-Hernández M., Lanzar Z., Lüdtke A., Perez O.A., Reizis B, & Idoyaga J. (2019). Integrated Cross-Species Analysis Identifies a Conserved Transitional Dendritic Cell Population. Cell reports, 29(11), 3736-3750.e8.
+ Open protocol
+ Expand
7

Mass Cytometry Analysis of Stimulated pDCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Metal-labeled Abs were obtained from Fluidigm or labeled using the MaxPar X8 labeling kit (Fluidigm) according to manufacturer’s instructions (see Table S2). For mass cytometry analysis of stimulated pDC experiments, bona fide pDCs (AXL−) were obtained by sorting as in Figure S3A. Freshly isolated and cultured pDCs were pooled with 3×106 mouse splenocytes to provide a cellular bed, stained for CyTOF, and identified as human CD45+ mouse CD45− cells. Cells were stained with 1 mL of 0.25 μM cisplatin (Fluidigm) for 5 minutes at room temperature to exclude dead cells. Cells were then washed with CyFACS buffer (2mM EDTA, 1% BSA 1% in PBS) and stained with heavy-metal-labeled Ab cocktail for 30 minutes on ice. Cells were washed twice with CyFACS then fixed with FoxP3 Transcription Factor Fix/Perm Buffer for 2 hours. Human surface Abs that were sensitive to FoxP3 buffer (i.e., CX3CR1, CD123, CD33, CD135, CD172a and CD163), were stained after fixation in 1X Permwash buffer, for 30 minutes at 4°C. After staining, samples were washed and incubated with 2% paraformaldehyde (Electron) in PBS containing 125 nM Iridium intercalator (Fluidigm) overnight. Cells were washed with water, filtered and acquired in a CyTOF2 (Fluidigm).
Leylek R., Alcántara-Hernández M., Granja J.M., Chavez M., Perez K., Diaz O.R., Li R., Satpathy A.T., Chang H.Y, & Idoyaga J. (2020). Chromatin Landscape Underpinning Human Dendritic Cell Heterogeneity. Cell reports, 32(12), 108180.
+ Open protocol
+ Expand
8

Mass Cytometry Analysis of Stimulated pDCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Metal-labeled Abs were obtained from Fluidigm or labeled using the MaxPar X8 labeling kit (Fluidigm) according to manufacturer’s instructions (see Table S2). For mass cytometry analysis of stimulated pDC experiments, bona fide pDCs (AXL−) were obtained by sorting as in Figure S3A. Freshly isolated and cultured pDCs were pooled with 3×106 mouse splenocytes to provide a cellular bed, stained for CyTOF, and identified as human CD45+ mouse CD45− cells. Cells were stained with 1 mL of 0.25 μM cisplatin (Fluidigm) for 5 minutes at room temperature to exclude dead cells. Cells were then washed with CyFACS buffer (2mM EDTA, 1% BSA 1% in PBS) and stained with heavy-metal-labeled Ab cocktail for 30 minutes on ice. Cells were washed twice with CyFACS then fixed with FoxP3 Transcription Factor Fix/Perm Buffer for 2 hours. Human surface Abs that were sensitive to FoxP3 buffer (i.e., CX3CR1, CD123, CD33, CD135, CD172a and CD163), were stained after fixation in 1X Permwash buffer, for 30 minutes at 4°C. After staining, samples were washed and incubated with 2% paraformaldehyde (Electron) in PBS containing 125 nM Iridium intercalator (Fluidigm) overnight. Cells were washed with water, filtered and acquired in a CyTOF2 (Fluidigm).
Leylek R., Alcántara-Hernández M., Granja J.M., Chavez M., Perez K., Diaz O.R., Li R., Satpathy A.T., Chang H.Y, & Idoyaga J. (2020). Chromatin Landscape Underpinning Human Dendritic Cell Heterogeneity. Cell reports, 32(12), 108180.
+ Open protocol
+ Expand
9

Multiparameter CyTOF Immunophenotyping

Check if the same lab product or an alternative is used in the 5 most similar protocols
Metal-labeled Abs (reporting summary) were obtained from Fluidigm or
labeled using the MaxPar X8 labeling kit (Fluidigm). CD135-enriched mouse cell
suspensions were stained with 0.25 μM cisplatin (Fluidigm) for 5 min at
RT to exclude dead cells, washed with CyFACS buffer (2 mM EDTA, 1% BSA, 1% in
PBS) and stained with a heavy-metal-labeled Abs for 30 min at 4°C. Cells
were washed twice with CyFACS then fixed with Foxp3 Transcription Factor
Fix/Perm Buffer for 2 hrs followed by intracellular staining in 1X Permwash for
30 min. After staining, samples were washed and incubated with 2% PFA in PBS
containing 125 nM Iridium intercalator (Fluidigm) overnight. Cells were washed
with water, filtered, and acquired. Human PBMC and sorted tDC were cisplatin and
surface stained, followed by fixation as described above for mouse. The
intracellular cocktail included Abs affected by fixation52 . Sorted tDC were pooled with mouse
splenocytes to avoid cell loss during washes52 . In all cases, cells were acquired in a CyTOF2 or a
Helios mass cytometer (Fluidigm) at the Shared FACS Facility at Stanford or Mt.
Sinai, using CyTOF software.
Sulczewski F.B., Maqueda-Alfaro R.A., Alcántara-Hernández M., Perez O.A., Saravanan S., Yun T.J., Seong D., Hornero R.A., Raquer-McKay H.M., Esteva E., Lanzar Z.R., Leylek R.A., Adams N.M., Das A., Rahman A.H., Gottfried-Blackmore A., Reizis B, & Idoyaga J. (2023). TRANSITIONAL DENDRITIC CELLS ARE DISTINCT FROM CONVENTIONAL DC2 PRECURSORS AND MEDIATE PRO-INFLAMMATORY ANTIVIRAL RESPONSES. Nature immunology, 24(8), 1265-1280.
+ Open protocol
+ Expand
10

Comprehensive Antibody Labeling Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
All anti-human pre-conjugated to metal isotopes were obtained from Fluidigm Corporation (San Francisco, US). All remaining antibodies were obtained from the indicated companies as purified antibodies and in-house conjugation was completed using the MaxPar X8 labeling kit (Fluidigm) following manufacturers’ recommended protocols. A detailed list of all antibodies is shown in the Key Resource table.
Souza-Moreira L., Tan Y., Wang Y., Wang J.P., Salkhordeh M., Virgo J., Florian M., Murray A.B., Watpool I., McIntyre L., English S., Stewart D.J, & Mei S.H. (2022). Poly(I:C) enhances mesenchymal stem cell control of myeloid cells from COVID-19 patients. iScience, 25(5), 104188.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!