Spectramax mz

Manufactured by Molecular Devices

The SpectraMax Mz is a multi-mode microplate reader designed for high-performance absorbance, fluorescence, and luminescence detection. It offers a broad wavelength range and flexible configuration options to support a variety of applications in life science research and drug discovery.

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Lab products found in correlation

Tissuelyser, by Qiagen (2 mentions) Trichloroacetic acid, by Merck Group (2 mentions) 5n naoh, by Merck Group (2 mentions) Biotinylated goat anti mouse igg, by Jackson ImmunoResearch (2 mentions) Tmb reagent set, by BD (2 mentions) Streptavidin horseradish peroxidase, by BD (2 mentions)

4 protocols using spectramax mz

Blood-Brain Barrier Permeability Assay

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BBB permeability was assessed using sodium-fluorescein (NaF) (Sigma), a low-molecular mass molecule (376 Da), to detect fluid-phase shifts between the circulation and central nervous system. Briefly, mice received 100 μl of 10% NaF in PBS intraperitoneally and cardiac blood was collected 10 min later. Mice were transcardially perfused with 10 ml of PBS, and spinal cord and brain removed. Each tissue was weighed, homogenized in PBS using a TissueLyser (Qiagen) and centrifuged at 14,000g for 2 min. A volume of 500 μl of clarified supernatant was added to 500 μl of 15% trichloroacetic acid (Sigma) and centrifuged at 10,000g for 10 min. A volume of 125 μl of 5 N NaOH (Sigma-Aldrich) was added to 500 μl of the supernatant and NaF content measured on a SpectraMax Mz (Molecular Devices) microplate reader using standards ranging from 125 to 4,000 μg. NaF in spinal cord or brain was normalized to sera NaF content using the formula: (mg fluorescence brain tissue per mg of protein)/(mg fluorescence sera per μl of blood). Data are expressed as fold increases in NaF in the spinal cord or brain with levels from WT-treated mice set at 1.

Kerr B.A., West X.Z., Kim Y.W., Zhao Y., Tischenko M., Cull R.M., Phares T.W., Peng X.D., Bernier-Latmani J., Petrova T.V., Adams R.H., Hay N., Naga Prasad S.V, & Byzova T.V. (2016). Stability and function of adult vasculature is sustained by Akt/Jagged1 signalling axis in endothelium. Nature Communications, 7, 10960.

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Virus Neutralizing Antibody Assay

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Virus neutralizing Ab was measured as described (39 (link)). Briefly, duplicates of serial 2-fold dilutions of heat inactivated serum from individual mice (n ≥ 5) were incubated with 50 PFU of JHMV in 96-well plates for 90 min at 37°C. Following addition of DBT cells (8 × 10⁴ cells/well) plates were incubated at 37°C for 48 hr. Neutralization titers represent the log₁₀ of the highest average serum dilution that inhibited cytopathic effect. Total virus-specific IgG was measured by ELISA as described (29 (link), 30 (link)). Briefly, plates were coated with 100 μl of a serum free supernatant derived from JMHV infected DBT cells and incubated overnight at 4°C. Plates were washed with PBS containing 0.05% Tween-20 and blocked with 10% FCS in PBS overnight at 4°C. Serum samples were diluted 2-fold in PBS and incubated overnight at 4°C. Following washing, bound Ab was detected using biotinylated goat anti-mouse IgG (Jackson ImmunoResearch Laboratories), streptavidin horseradish peroxidase (BD Bioscience) and 3,3′,5,5′ Tetramethylbenzidine (TMB Reagent Set; BD Bioscience). Optical densities were read at 450 nm on a SpectraMax Mz (Molecular Devices, Sunnydale, CA) microplate reader and titers calculated as reciprocals of the highest dilution that exceeded 2-fold ± standard errors of the means (SEM) over naive controls.

Phares T.W., DiSano K.D., Stohlman S.A., Segal B.M, & Bergmann C.C. (2016). CXCL13 promotes isotype-switched B cell accumulation to the central nervous system during viral encephalomyelitis. Brain, Behavior, and Immunity, 54, 128-139.

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Virus Neutralization and Total IgG Assays

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Virus neutralizing Ab was measured as described (Tschen et al., 2002 (link)). Briefly, duplicates of serial 2-fold dilutions of heat inactivated serum from individual mice (n ⩾ 5) were incubated with 50 PFU of JHMV in 96-well plates for 90 min at 37 °C. Following addition of DBT cells (8 × 10⁴ cells/well) plates were incubated at 37 °C for 48 h. Neutralization titers represent the log₁₀ of the highest average serum dilution that inhibited cytopathic effect. Total virus-specific IgG was measured by ELISA as described (Tschen et al., 2002 (link), Tschen et al., 2006 (link)). Briefly, plates were coated with 100 μl of a serum free supernatant derived from JMHV infected DBT cells and incubated overnight at 4 °C. Plates were washed with PBS containing 0.05% Tween-20 and blocked with 10% FCS in PBS overnight at 4 °C. Serum samples were diluted 2-fold in PBS and incubated overnight at 4 °C. Following washing, bound Ab was detected using biotinylated goat anti-mouse IgG (Jackson ImmunoResearch Laboratories), streptavidin horseradish peroxidase (BD Bioscience) and 3,3′,5,5′ Tetramethylbenzidine (TMB Reagent Set; BD Bioscience). Optical densities were read at 450 nm on a SpectraMax Mz (Molecular Devices, Sunnydale, CA) microplate reader and titers calculated as reciprocals of the highest dilution that exceeded 2-fold ± standard errors of the means (SEM) over naive controls.

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Assessing Blood-Brain Barrier Permeability

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To detect fluid-phase shifts between the circulation and CNS, BBB permeability
was assessed using sodium-fluorescein (NaF). Briefly, mice received 100 μL of
10% NaF in PBS (IP) and cardiac blood was collected 10 min later. Mice were
transcardially perfused with 10 mL of PBS and brains were removed. Each tissue
was weighed, homogenized in PBS using a TissueLyser (Qiagen), and centrifuged at
14,000 g for 2 min. A volume of 500 μL of clarified
supernatant was added to 500 μL of 15% trichloroacetic acid (Sigma) and
centrifuged at 10,000 g for 10 min. A volume of 125 μL of 5 N
NaOH (Sigma-Aldrich) was added to 500 μL of the supernatant and NaF content was
measured on a SpectraMax Mz (Molecular Devices) microplate reader using
standards ranging from 125 to 4,000 μg. NaF in the brain was normalized to sera
NaF content using the formula: (μg of NaF in brain tissue per mg of protein)/(μg
of NaF in sera per μL of blood). Data are expressed as fold increases in NaF in
the brain, with levels from PBS-treated mice set at 1. For the positive control,
C57BL/6 mice were infected at 6 to 7 weeks of age by intracranial injection into
the left cerebral hemisphere with 1,000 plague-forming units (PFU) of the
J.2.2v-1 monoclonal Ab-derived glia-tropic JHMV variant in 30 μL sterile PBS
(Fleming et al.,
1986; Phares et
al., 2014).

Chen Z., Hu W., Mendez M.J., Gossman Z.C., Chomyk A., Boylan B.T., Kidd G.J., Phares T.W., Bergmann C.C, & Trapp B.D. (2023). Neuroprotection by Preconditioning in Mice is Dependent on MyD88-Mediated CXCL10 Expression in Endothelial Cells. ASN NEURO, 15, 17590914221146365.

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