Sector s 600

Manufactured by MSD

The Sector S 600 is a versatile laboratory equipment designed for a wide range of applications. It is a compact and efficient device that performs essential tasks in research and analytical settings. The core function of the Sector S 600 is to provide reliable and consistent results, enabling users to conduct their experiments and analyses with confidence.

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Lab products found in correlation

Amicon ultra centrifugal filter device, by Merck Group (1 mentions) Blocker a, by MSD (1 mentions) Diluent 100, by MSD (1 mentions) Facs lyse solution, by BD (1 mentions) Navios flow cytometer, by Beckman Coulter (1 mentions) Duoset elisa kit, by R&D Systems (1 mentions) V plex proinflammatory panel 1 nhp kit, by Mesoscale (1 mentions) Anti human igm, by Jackson ImmunoResearch (1 mentions) Phosstop, by Roche (1 mentions) Complete mini, by Roche (1 mentions)

9 protocols using sector s 600

SARS-CoV-2 Variant S-2P Binding Inhibition Assay

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ACE2 binding inhibition was performed using a modified MSD platform assay. Briefly, after blocking MSD Streptavidin MULTI-ARRAY 384 well plates with Blocker A (MSD), the plates were coated with 1 μg/ml of biotinylated SARS-CoV-2 variant S-2P (D614G, Beta, Delta, or Omicron) and incubated for 1 hour at room temperature (RT). The plates were washed 5 times with wash buffer (1x PBS containing 0.05% Tween-20). Diluted samples were added to the coated plates and incubated for 1 hour at RT. MSD SULFO-TAG human ACE2 protein was diluted 1:200 and added to the plates. After 1 hour incubation at RT, the plates were washed 5 times with wash buffer and read on MSD Sector S 600 instrument after the addition of Gold Read Buffer B (MSD). Results are reported as percent inhibition. BAL fluid and nasal washes were first concentrated 10-fold with Amicon Ultra centrifugal filter devices (Millipore Sigma) and then diluted 1:5 in Diluent 100 (MSD).

Gagne M., Moliva J.I., Foulds K.E., Andrew S.F., Flynn B.J., Werner A.P., Wagner D.A., Teng I.T., Lin B.C., Moore C., Jean-Baptiste N., Carroll R., Foster S.L., Patel M., Ellis M., Edara V.V., Maldonado N.V., Minai M., McCormick L., Honeycutt C.C., Nagata B.M., Bock K.W., Dulan C.N., Cordon J., Flebbe D.R., Todd J.P., McCarthy E., Pessaint L., Van Ry A., Narvaez B., Valentin D., Cook A., Dodson A., Steingrebe K., Nurmukhambetova S.T., Godbole S., Henry A.R., Laboune F., Roberts-Torres J., Lorang C.G., Amin S., Trost J., Naisan M., Basappa M., Willis J., Wang L., Shi W., Doria-Rose N.A., Zhang Y., Yang E.S., Leung K., O’Dell S., Schmidt S.D., Olia A.S., Liu C., Harris D.R., Chuang G.Y., Stewart-Jones G., Renzi I., Lai Y.T., Malinowski A., Wu K., Mascola J.R., Carfi A., Kwong P.D., Edwards D.K., Lewis M.G., Andersen H., Corbett K.S., Nason M.C., McDermott A.B., Suthar M.S., Moore I.N., Roederer M., Sullivan N.J., Douek D.C, & Seder R.A. (2022). mRNA-1273 or mRNA-Omicron boost in vaccinated macaques elicits similar B cell expansion, neutralizing responses, and protection from Omicron. Cell, 185(9), 1556-1571.e18.

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STING1 KO HMC3 Cell Cytokine Assay

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Following 72 h pre-treatment with 10 nM AZD1390 or DMSO as a control, wild-type (WT) and STING1 KO HMC3 cells were seeded at a density of 21 000 cells/cm² in 6-well plates. After 48 h, cells were washed in PBS and placed in DMEM supplemented with 0.5% (v/v) HI-FBS. Cells were stimulated with 1 μM diABZi for 24 h prior to collection. Conditioned medium was centrifuged at 400 g for 10 min at 4ºC to pellet cells and debris, then snap frozen on dry ice and stored at -80ºC until use. After collection of medium, remaining cells were lysed in RIPA buffer and total protein content was measured by Bradford assay. The overall protein content was comparable between treatments.
Secretion of IFN-α2a, IFN-β and CXCL10 was measured using a Mesoscale Discovery (MSD) U-PLEX assay following the manufacturer's protocol. Plates were read on a MESO SECTOR S 600 (MSD), and data was analysed using MSD Discovery Workbench (version 4).

Talbot E.J., Joshi L., Thornton P., Dezfouli M., Tsafou K., Perkinton M, & Khoronenkova S.V. (2023). cGAS-STING signalling regulates microglial chemotaxis in genome instability. Nucleic Acids Research, 52(3), 1188-1206.

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Quantification of Anti-CTLA-4 and Anti-PD-1 Antibodies

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The respective antigen (CTLA-4 or PD-1 protein) was diluted into PBS and coated onto each well of a microtiter plate for at least 12 hours at 4°C. Plates were washed 5-fold with 25mM Tris, 150 mM NaCl, 0.1% Tween 20, pH 7.5 (wash buffer). 150 μl of blocking assay buffer (Superblock TBS-T20, Thermo Fisher Scientific) was added, and the plates were sealed and incubated for at least 1 hour with constant shaking. Plasma samples collected pre- and 5 mins post-infusion were serially diluted 50-fold into assay buffer. Samples needing further dilution were diluted into assay buffer containing 2% cynomolgus macaque serum. After washing, sample was added to each well of the microtiter plate as appropriate: test sample, blank, calibration standard, or quality controls (i.e antibody diluted into plasma from healthy RMs, Bioreclamation). Plates were sealed and incubated with constant shaking for 2 hours at 37°C. Plates were then washed, detection solution added (ruthenium-labelled mouse anti-human IgG Fc JDC-10 from Southern Biotech or ruthenium-labelled PD-1 from GSK), and plates were incubated with constant shaking for 1 hour at 37°C. Plates were once again washed, read buffer T (Meso Scale Discovery) was added to each well, and plates were read within 5 mins on an MSD Sector S600.

Harper J., Gordon S., Chan C.N., Wang H., Lindemuth E., Galardi C., Falcinelli S.D., Raines S.L., Read J.L., Nguyen K., McGary C.S., Nekorchuk M., Busman-Sahay K., Schawalder J., King C., Pino M., Micci L., Cervasi B., Jean S., Sanderson A., Johns B., Koblansky A.A., Amrine-Madsen H., Lifson J., Margolis D.M., Silvestri G., Bar K.J., Favre D., Estes J.D, & Paiardini M. (2020). CTLA-4 and PD-1 dual blockade induces SIV reactivation without control of rebound after ART interruption. Nature medicine, 26(4), 519-528.

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Soluble CD137 and Cell Subsets Quantification

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Soluble CD137/4-1BB (sCD137/4-1BB) in plasma was measured using a customized sandwich assay with goat anti-human 4-1BB capture antibody and a biotinylated goat anti-human 4-1BB detection antibody from the human 4-1BB/TNFRSF9 Duo Set ELISA kit (R&D Systems, DY838). This assay was run on a Meso Scale Discovery (MSD) platform.
The MSD standard 96-well plates were pre-coated with the goat antihuman 4-1BB capture antibody. After incubation with the plasma samples, biotinylated goat anti-human 4-1BB detection antibody was added to the plates, then SULFO-TAG labeled Streptavidin (MSD, K15A01-1) was added to the detection antibody. Finally, an MSD read buffer was added and the plates were subject to electrochemiluminescence signal reading on an MSD SECTOR S600. Soluble CD137 concentrations in the plasma samples were then quantified based on the standard calibration curve present on the same plate.
For the determination of B cells, NK cells and T cells, we used a BECKMAN COULTER NAVIOS flow cytometer. A portion of the blood was added to the staining tubes, which contained BD MultiTEST CD3-FITC/CD16&CD56-PE/CD45-PerCP-Cv5.5/CD4-PE-Cy7/CD19-APC/CD8-APC-Cy7 (BD Biosciences) and calibrated counting beads. After lysing red blood cells with FACSLyse solution (BD Biosciences), we washed and fixed the specimens with 1% paraformaldehyde solution and stored them at 4°C until acquisition.

Ma Y., Luo F., Zhang Y., Liu Q., Xue J., Huang Y., Zhao Y., Yang Y., Fang W., Zhou T., Chen G., Cao J., Chen Q., She X., Luo P., Liu G., Zhang L, & Zhao H. (2024). Preclinical characterization and phase 1 results of ADG106 in patients with advanced solid tumors and non-Hodgkin’s lymphoma. Cell Reports Medicine, 5(2), 101414.

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Plasma PK and Cytokine Profiling Workflow

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Whole blood from ACD tubes was centrifuged to obtain plasma for PK and cytokine profiling. GSK445A concentration was determined from plasma by LC-MS/MS analysis (Frontage). Plasma concentrations of NHP IL-6 was determined according the manufacturer’s recommendations for the V-PLEX Proinflammatory Panel 1 NHP Kit (Meso Scale Discovery) using a 1:1 dilution of NHP plasma and Diluent 2 plus 1% Triton-X100. IL-6 was measured on a Meso Scale Discovery Sector S 600 plate reader and analysis carried out with MSD Discovery Workbench v4.0.12 and GraphPad PRISM 6.

Okoye A.A., Fromentin R., Takata H., Brehm J.H., Fukazawa Y., Randall B., Pardons M., Tai V., Tang J., Smedley J., Axthelm M., Lifson J.D., Picker L.J., Favre D., Trautmann L, & Chomont N. (2022). The ingenol-based protein kinase C agonist GSK445A is a potent inducer of HIV and SIV RNA transcription. PLoS Pathogens, 18(1), e1010245.

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Fractalkine and CX3CR1 in EAC

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The V-PLEX™ Fractalkine plate (Meso Scale Discovery) was used to detect the levels of fractalkine in the serum and ACM of 19 EAC patients according to the manufacturer’s instructions and read using an MSD Sector S600. The Human CX3CR1 ELISA Kit (ELISA Genie) was used to measure secreted CX3CR1 in the serum and ACM of 10 EAC patients and read using a VersaMax™ ELISA microplate reader.

Conroy M.J., Maher S.G., Melo A.M., Doyle S.L., Foley E., Reynolds J.V., Long A, & Lysaght J. (2018). Identifying a Novel Role for Fractalkine (CX3CL1) in Memory CD8+ T Cell Accumulation in the Omentum of Obesity-Associated Cancer Patients. Frontiers in Immunology, 9, 1867.

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Evaluating SARS-CoV-2 Neutralizing Antibodies

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Meso Scale Discovery (MSD) V-PLEX SARS-CoV-2 ACE2 Neutralization Kit, Panels 5 and 14 were used to evaluate sera collected from immunized study animals according to the manufacturer’s instructions with the MSD Sector S 600 instrument. Briefly, MSD plates containing SARS-CoV-2 Spike proteins (wildtype, Beta, Gamma and Delta) were blocked, washed, and incubated with sera from vaccinated animals at a 1:27 dilution. Plates were then washed and incubated with SULFO-TAG ACE2 and developed according to the manufacturer’s protocol. Functional antibody activity was measured as percent inhibition of binding of SULFO-TAG ACE2 to Spike protein.

Walters J.N., Schouest B., Patel A., Reuschel E.L., Schultheis K., Parzych E., Maricic I., Gary E.N., Purwar M., Andrade V.M., Doan A., Elwood D., Eblimit Z., Nguyen B., Frase D., Zaidi F.I., Kulkarni A., Generotti A., Joseph Kim J., Humeau L.M., Ramos S.J., Smith T.R., Weiner D.B, & Broderick K.E. (2022). Prime-boost vaccination regimens with INO-4800 and INO-4802 augment and broaden immune responses against SARS-CoV-2 in nonhuman primates. Vaccine, 40(21), 2960-2969.

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Cell-Based Binding Assay for SD

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ECL cell-based binding assays were performed using MSD MULTI-ARRAY 96 HB plates (hereafter referred to as "high bind plates"). For titration of SD, CD47 CHO-K1 cells were collected from continuous culture and incubated with SD in V-bottom polypropylene (PP) plates to allow binding of drug to its target and to facilitate removal of unbound drug by washing cells with PBS. Cells were washed twice by resuspension with 200-250 µL PBS per well, centrifugation at 300 × g, and aspiration of PBS. Washed cells were transferred to MSD high bind plates in PBS and allowed to settle/attach. MSD Read Buffer T surfactant-free was added to each well, and the high bind plates were read on an MSD Sector S 600 instrument. For sensitivity, DT, and interference assessments, CD47 CHO-K1 cells were incubated with a 1:1 mixture of SD and sample (i.e., product of sample pretreatment).

Luong M., Wang Y., Donnelly B.B, & Lepsy C. (2023). Addressing Domain Specificity in the Development of a Cell-Based Binding Assay for the Detection of Neutralizing Antibodies Against a CD47xPD-L1 Bispecific Antibody. The AAPS journal, 25(6).

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Quantification of PLCγ2 Phosphorylation

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Ramos cells (ATCC, Manassas, VA) were treated with titrating concentrations of BIIB091 for 30 min at 37°C in duplicates and subsequently stimulated for 5 min with anti‐human IgM (1 µg mL⁻¹; Jackson ImmunoResearch, West Grove, PA). After centrifugation and removal of the supernatant, the cell pellets were lysed with gentle shaking in cold lysis buffer (Promega, Madison, WI) containing protease (Complete Mini; Roche, Basel, Switzerland) and phosphatase (PhosSTOP™, Roche); and phosphatase inhibition cocktail 2 and 3 (Sigma‐Aldrich) inhibitors. Cell lysates (diluted) were added and incubated for approximately 12 h at 4°C in MSD plate coated with anti‐PLCγ2 antibody (diluted 1:100, B‐10; Santa Cruz Biotechnology, Dallas, TX). Phosphorylated PLCγ2 was detected with rabbit polyclonal anti‐phospho tyrosine 1217 of PLCγ2 (diluted 1:1000; Cell Signaling Technologies) and Sulfo‐Tag goat anti‐rabbit antibody (diluted 1:500; MSD). Levels of phosphorylated PLCγ2 were detected as ECL signal on a Sector S600 (MSD) after the addition of 1× MSD Read Buffer.

Bame E., Tang H., Burns J.C., Arefayene M., Michelsen K., Ma B., Marx I., Prince R., Roach A.M., Poreci U., Donaldson D., Cullen P., Casey F., Zhu J., Carlile T.M., Sangurdekar D., Zhang B., Trapa P., Santoro J., Muragan P., Pellerin A., Rubino S., Gianni D., Bajrami B., Peng X., Coppell A., Riester K., Belachew S., Mehta D., Palte M., Hopkins B.T., Scaramozza M., Franchimont N, & Mingueneau M. (2021). Next‐generation Bruton's tyrosine kinase inhibitor BIIB091 selectively and potently inhibits B cell and Fc receptor signaling and downstream functions in B cells and myeloid cells. Clinical & Translational Immunology, 10(6), e1295.

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