The uronic acid content of the PE fraction was measured according to Blumenkrantz and Asboe-Hansen (1973) . Briefly, 100 μl extracts were incubated in glass tubes with 500 μl 98% H 2 SO 4 containing 0.0125 M Na 2 B 4 O 7 ⋅10 H 2 O for 5 min at 100 ℃. After cooling, 10 μl of m-Hydroxydiphenyl (0.15%) was added and after 20 min incubation at room temperature, the absorbance at 520 nm was measured using the BioTek Gen5 Plate Reader (BioTek, USA). Galacturonic acid (Sigma) was used to construct the calibration curve. Total polysaccharide contents of HC1 and HC2 fractions were measured according to the phenol sulfuric acid method of Dubois et al. (1956) and Shi et al. (2015) . Briefly, 100 μl extracts were firstly incubated with 500 μl 98% H 2 SO 4 and 5 μl 80% phenol in glass tubes at room temperature for 15 min, then were incubated at 100 ℃ for another 15 min. After cooling, the absorbance at nm was measured by the BioTek Gen5 Plate Reader (BioTek, USA). Glucose was used to construct the calibration curve.
Biotek gen5 plate reader
Manufactured by Agilent Technologies
Sourced in United States
The BioTek Gen5 plate reader is a versatile instrument designed for a wide range of microplate-based applications. It offers absorbance, fluorescence, and luminescence detection capabilities, enabling researchers to perform various types of assays, including cell-based, biochemical, and molecular biology experiments.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated goat anti mouse secondary antibody, by Jackson ImmunoResearch (2 mentions)
Sample buffer, by RayBiotech (2 mentions)
Anti rabbit antibodies, by Jackson ImmunoResearch (2 mentions)
Ieq cov s rbd igg, by RayBiotech (2 mentions)
Tmb substrate, by RayBiotech (2 mentions)
Ieq cov n igg, by RayBiotech (2 mentions)
Galacturonic acid, by Merck Group (1 mentions)
3 protocols using biotek gen5 plate reader
1
Quantifying Uronic Acids and Polysaccharides
2
SARS-CoV-2 Antibody Detection by ELISA
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Mouse and rabbit IgG antibodies against SARS-CoV-2 spike or Ncap were measured by enzyme-linked immunosorbent assays (ELISAs) using precoated ELISA plates (IEQ-CoV-S-RBD-IgG and IEQ-CoV-N-IgG; RayBiotech), according to the manufacturer’s instructions, at RT. Briefly, plasma samples were diluted in sample buffer (RayBiotech), added to antigen-coated wells in triplicates, and incubated at RT for 2 h on a shaker (200 rpm). Commercially available antibody against spike (catalog number S1N-S58; Acro Biosystems) or Ncap (catalog number NUN-S47; Acro Biosystems) was used as a positive control. Plates were washed 3 times with wash buffer and incubated for 1 h at RT with HRP-conjugated goat anti-mouse secondary antibodies (dilution of 1:5,000) (catalog number 115-035-003; Jackson ImmunoResearch) or anti-rabbit antibodies (dilution of 1:5,000) (catalog number 111-035-003; Jackson ImmunoResearch) diluted in assay buffer (RayBiotech). After 3 washes, the plates were developed using the TMB substrate (RayBiotech). After a 15-min incubation, the reaction was stopped by the addition of a stop solution, and the absorbance at 450 nm was recorded using a BioTek Gen5 plate reader (Agilent). Endpoint titers were calculated as the dilution that emitted an optical density (OD) exceeding 4 times that for the PBS control group.
3
SARS-CoV-2 Antibody Detection by ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse and rabbit IgG antibodies against SARS-CoV-2 spike or Ncap were measured by enzyme-linked immunosorbent assays (ELISAs) using precoated ELISA plates (IEQ-CoV-S-RBD-IgG and IEQ-CoV-N-IgG; RayBiotech), according to the manufacturer’s instructions, at RT. Briefly, plasma samples were diluted in sample buffer (RayBiotech), added to antigen-coated wells in triplicates, and incubated at RT for 2 h on a shaker (200 rpm). Commercially available antibody against spike (catalog number S1N-S58; Acro Biosystems) or Ncap (catalog number NUN-S47; Acro Biosystems) was used as a positive control. Plates were washed 3 times with wash buffer and incubated for 1 h at RT with HRP-conjugated goat anti-mouse secondary antibodies (dilution of 1:5,000) (catalog number 115-035-003; Jackson ImmunoResearch) or anti-rabbit antibodies (dilution of 1:5,000) (catalog number 111-035-003; Jackson ImmunoResearch) diluted in assay buffer (RayBiotech). After 3 washes, the plates were developed using the TMB substrate (RayBiotech). After a 15-min incubation, the reaction was stopped by the addition of a stop solution, and the absorbance at 450 nm was recorded using a BioTek Gen5 plate reader (Agilent). Endpoint titers were calculated as the dilution that emitted an optical density (OD) exceeding 4 times that for the PBS control group.
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