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Spectramax m5

Manufactured by PerkinElmer
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The SpectraMax M5 is a multi-mode microplate reader from PerkinElmer. It is designed to perform a variety of absorbance, fluorescence, and luminescence assays. The instrument can be used to measure various biological and chemical samples in microplate format.

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Lab products found in correlation

Quantification kits, by Nanjing Jiancheng (1 mentions) Envision microplate reader, by PerkinElmer (1 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions) Suc llvy amc, by Enzo Life Sciences (1 mentions) 96 well plate, by USA Scientific (1 mentions) Blocking buffer, by LI COR (1 mentions) Ha tag rabbit monoclonal antibody, by Cell Signaling Technology (1 mentions) Streptavidin hrp, by PerkinElmer (1 mentions)

5 protocols using spectramax m5

1

Quantification Kit-Based Absorbance Assay

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All samples were measured using quantification kits (Jiancheng Bioengineering, Nanjing, China) according to the manufacturer’s instructions, and then absorbance was determined by EnVision Microplate reader (Molecular Devices SpectraMax M5, PerkinElmer, USA).
Cheng L., Ge M., Lan Z., Ma Z., Chi W., Kuang W., Sun K., Zhao X., Liu Y., Feng Y., Huang Y., Luo M., Li L., Zhang B., Hu X., Xu L., Liu X., Huo Y., Deng H., Yang J., Xi Q., Zhang Y., Siegenthaler J.A, & Chen L. (2017). Zoledronate dysregulates fatty acid metabolism in renal tubular epithelial cells to induce nephrotoxicity. Archives of Toxicology, 92(1), 469-485.
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2

Zoledronate-Induced Cell Viability Assay

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Cell viability was measured by using MTT assay. Briefly, HK-2 cells were seeded into a 96-well plate and exposed to zoledronate (0, 0.1, 1, 5, 10, 50 µM). After treatment for varying periods of time (24, 36, 48, 60, or 72 h), 20 μl MTT at 5 mg/ml was added to each well. The cells were incubated at 37 °C for another 4 h and then dimethyl sulfoxide (DMSO) (Thermo Fisher Scientific, USA) was added to each well. The absorbance was detected at 570 nm with a microplate reader (Molecular Devices SpectraMax M5, PerkinElmer, USA). Cell viability was expressed as a percentage of the control culture.
Cheng L., Ge M., Lan Z., Ma Z., Chi W., Kuang W., Sun K., Zhao X., Liu Y., Feng Y., Huang Y., Luo M., Li L., Zhang B., Hu X., Xu L., Liu X., Huo Y., Deng H., Yang J., Xi Q., Zhang Y., Siegenthaler J.A, & Chen L. (2017). Zoledronate dysregulates fatty acid metabolism in renal tubular epithelial cells to induce nephrotoxicity. Archives of Toxicology, 92(1), 469-485.
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3

Proteasome Inhibition Assay Protocol

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Human recombinant 20S proteasome (Enzo Life Sciences; Farmingdale, NY) was assayed using fluorogenic substrates. The chymotrypsin-like, trypsin-like, and caspase-like sites were independently assayed using the selective substrates Suc-LLVY-AMC, Cbz-LLE-AMC, and Cbz-VGR-AMC, respectively (Enzo Life Sciences; Farmingdale, NY). An 11-point, 3-fold serial dilution dose-response assay was performed in triplicate. Inhibitor (10 μL) or vehicle control in 25% DMSO was added to each well of a 96-well plate, followed by 30 μL of assay buffer (100 mM Tris, pH 7.5, EDTA 0.5 mM, SDS 0.03%) containing 3 μg/mL of 20S proteasome. After 30 minutes of incubation at 37°C, 10 μL of 200 μM substrate was added to initiate proteasome reaction, which proceeded for 30 min at 37°C. The resultant dose response concentration range was 1.0 μM to 0.17 nM inhibitor in a 50 μL final reaction volume. The plates were analyzed on a SpecraMax Gemini or SpectraMax M5 microplate reader (PerkinElmer Life Sciences) and the fluorescent signal was measured at the excitation and emission wavelengths of 365 and 450 nm, respectively. Data were scaled to internal controls and a four-parameter logistic model (GraphPad vs. 5.0, Prism) was used to fit the measured data and determine IC50 values.
Almaliti J., Miller B., Pietraszkiewicz H., Glukhov E., Benjamin Naman C., Kline T., Hanson J., Li X., Zhou S., Valeriote F.A, & Gerwick W.H. (2018). Exploration of the Carmaphycins as Payloads in Antibody Drug Conjugate Anticancer Agents. European journal of medicinal chemistry, 161, 416-432.
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4

Modified Tat ELISA for Protein Detection

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Since we couldn’t find a good commercial Tat ELISA kit. We used a modified Tat ELISA protocol. Briefly, 96-well plate (USA Scientific) was coated with 0.2 μg HA-tag rabbit monoclonal antibody (Cell Signaling) per well. Wash the wells with TBST and block the wells with blocking buffer (Licor) for 1 h at room temperature. Add 100 μL sample to specific wells and incubate at 4 °C overnight. After washing with TBST, Anti-HIV1 tat antibody (Biotin) (Abcam) in blocking buffer with 1:100 dilution was added to each well and incubate at room temperature for 2 h. Wash the wells with TBST for three times. Incubate with 100 μl diluted Streptavidin-HRP (1:100 dilution) (PerkinElmer) for 30 min at room temperature. After washing with TBST for 3 times, incubate with 100 μl OPD (O-Phenylenediamine Dihydrochloride) (PerkinElmer) for 30 min at room temperature, add 100 μl stop solution (PerkinElmer) to terminate the reaction, measure the absorbance at wavelength 490 nm using SpectraMax M5.
Tang X., Lu H, & Ramratnam B. (2020). Neurotoxicity of HIV-1 Tat is attributed to its penetrating property. Scientific Reports, 10, 14002.
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5

Bacterial Growth Quantification Methods

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Growth was measured by diluting sample, plating on LB agar, and enumeration of resulting colonies or by measuring absorbances (600 nm) of population subsamples in a Beckman Coulter spectrophotometer (Model DU730 A23616, 1 cm path length) and in PerkinElmer EnVision or Spectramax M5 plate-readers.
Bruger E.L., Synder D.J., Cooper V.S., & Waters C.M. (2020). Quorum sensing provides a molecular mechanism for evolution to tune and maintain investment in cooperation.
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