S1599

Manufactured by Agilent Technologies

The S1599 is a high-performance benchtop Ultra-High-Performance Liquid Chromatography (UHPLC) system designed for analytical and preparative chromatography applications. It offers precise and reliable solvent delivery, automatic sample handling, and advanced data analysis capabilities to support a wide range of laboratory workflows.

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Lab products found in correlation

Nunc immobilizer, by Thermo Fisher Scientific (2 mentions) Peroxidase affinipure goat anti human igg h l, by Jackson ImmunoResearch (1 mentions) 96 well elisa plate, by Corning (1 mentions) Multiskan spectrum, by Thermo Fisher Scientific (1 mentions) Tween 20, by Merck Group (1 mentions) Lb509, by Abcam (1 mentions) Omega plate reader, by Agilent Technologies (1 mentions)

4 protocols using s1599

ELISA for Detecting PCNA-NKp44 Interaction

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A 96-well ELISA plate (Costar) was coated overnight at 4°C with
0.1μM of the recombinant human PCNA (His and MBP tagged) and 0.1μM
of His-DJ1 (Human, negative control, a kind gift from Dr. Dan Levy, BGU, Israel)
in PBS. After washing, the plate was blocked with 2.5% skimmed milk in PBST
(PBS+0.05% TWEEN-20 [Sigma, P1379]) for 1h at 37°C followed by washing
and incubation with 2.5μg/ml of 14-25-9 and purified mouse IgG1κ
isotype control (BioLegend, 401402). Detection was done using mouse IgG
HRP-linked whole Ab (from Sheep) (GE Healthcare, NA931) (1:1000 final
dilutions). Following washing, TMB (DAKO, S1599) was added. Optical density
(O.D.) was measured at 650 nm (Thermo Electron Multiskan Spectrum).
In another setup, ELISA plates were coated overnight at 4°C with
0.1μM of the recombinant human PCNA (MBP tagged); blocking was done as
above. After blocking the plates were washed and incubated with 10 to
0.65μg/ml of 14-25-9 and PC10, 2.5μg/ml of mouse IgG1 and 0.25%
skimmed milk in PBST as control for 1h at 37°C and then NKp44-Ig (2
μg/ml, final concentration) was added without washing for 1h at
37°C. NKp44-Ig binding to PCNA was detected using peroxidase AffiniPure
goat anti-human IgG (H+L) (Jackson ImmunoResearch Laboratories,
Inc.) (1:1000 final dilution). Following washing, TMB
(100μl/well DAKO, S1599) was added. O.D. was read at 650 nm (Thermo
Electron Multiskan Spectrum).

Kundu K., Ghosh S., Sarkar R., Edri A., Brusilovsky M., Gershoni-Yahalom O., Yossef R., Shemesh A., Soria J.C., Lazar V., Ben-Zion J., Campbell K.S., Elkabets M, & Porgador A. (2019). Inhibition of the NKp44-PCNA Immune Checkpoint Using a mAb to PCNA. Cancer immunology research, 7(7), 1120-1134.

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Quantifying Alpha-Synuclein and Tau Oligomers

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ELISA plates were coated with 1 µg/well of α-Syn monomer, α-Syn oligomeric polymorphs, and tau oligomer. 0.1 M sodium bicarbonate, pH 9.6, was used as a coating buffer followed by overnight incubation with primary antibodies: SynTC1 (1:4000), SynTC2 (1:1000), SynTC3 (1:1000), and total α-Syn commercial antibody LB509 (1:5000; Abcam 27,766) at 4 °C. Plates were then washed three times with TBS-T and incubated with 100 μl of HRP-conjugated anti-mouse IgG, diluted in 5% nonfat milk in TBS-T, for 1 h at room temperature. Plates were washed three times with TBS-T and developed with 3,3,5,5-tetramethylbenzidine (S1599, Dako). The reaction was stopped using 100 μl of 1 m HCl, and absorbance was read at 450 nm using a POLARstar OMEGA plate reader. All experiments were performed in triplicate [42 (link), 43 (link)].

Moore K., Sengupta U., Puangmalai N., Bhatt N, & Kayed R. (2023). Polymorphic Alpha-Synuclein Oligomers: Characterization and Differential Detection with Novel Corresponding Antibodies. Molecular Neurobiology, 60(5), 2691-2705.

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Quantifying Anti-Synuclein Antibody Responses

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Anti-syn oligomer antibody response was determined by screening serial dilutions of animal sera using an Indirect enzyme-linked immunosorbent assay (ELISA) as previously published [42 (link), 43 (link)]. Briefly, 96-well plates (Nunc Immobilizer, Amino Plates and Modules, 436,006, Thermo Fisher Scientific) were previously coated with 1 μl α-Syn oligomers, Aβ oligomers, or tau oligomers in 50 μl of 1 × PBS, pH 7.4, as coating buffer. After washing three times with TBS-T, plates were blocked for 2 h at room temperature with 10% nonfat milk in TBS-T. Plates were then washed three times with TBS-T and probed with 100 μl of primary antibodies for 1 h at room temperature (RT): commercial antibodies, LB509 (1:5000;Abcam 27,766), Syn211 (1:5000;Abcam 80,627)), sequence-specific α-Syn antibodies Syn 33 (1:1000;SigmaAldrich ABN2265), MJFR (1:1000;Abcam 209,538), and F8H7 (1:1000). Plates were then washed three times with TBS-T and incubated with 100 μl of HRP-conjugated anti-rabbit or anti-mouse IgG, diluted 1:10,000 in 5% nonfat milk in TBS-T, for 1 h at room temperature. Plates were washed three times with TBS-T and developed with 3,3,5,5-tetramethylbenzidine (S1599, Dako). The reaction was stopped using 100 μl of 1 m HCl, and absorbance was read at 450 nm using a POLARstar OMEGA plate reader. All experiments were performed in triplicate.

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ELISA Assay for Tau Protein

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An ELISA was conducted as described previously (24 (link)). Briefly, 96-well plates (Nunc Immobilizer, Amino Plates and Modules, 436006, Thermo Fisher Scientific) were previously coated with 1.5 μl of BDTOs in the presence or absence of curcumin derivatives using 50 μl of 1× PBS, pH 7.4, as coating buffer. After washing three times with TBS-T, plates were blocked for 2 h at room temperature with 120 μl of 10% nonfat milk in TBS-T. Plates were then washed three times with TBS-T and probed with 100 μl of primary antibodies for 1 h at room temperature (T22 (1:250) and T18 (1:2000)) and sequence-specific tau antibodies (Tau 13 (1:2000), Tau 5 (1:2000), Tau1 (1:2000), Tau 46 (1:2000), HT7 (1:2000), and 4G8 (1:1000)). Plates were then washed three times with TBS-T and incubated with 100 μl of HRP-conjugated anti-rabbit or anti-mouse IgG, diluted 1:10,000 in 5% nonfat milk in TBS-T, for 1 h at room temperature. Plates were washed three times with TBS-T and developed with 3,3,5,5-tetramethylbenzidine (S1599, Dako) The reaction was stopped using 100 μl of 1 m HCl, and absorbance was read at 450 nm using a POLARstar OMEGA plate reader. All experiments were performed in triplicate.

Lo Cascio F., Garcia S., Montalbano M., Puangmalai N., McAllen S., Pace A., Palumbo Piccionello A, & Kayed R. (2020). Modulating disease-relevant tau oligomeric strains by small molecules. The Journal of Biological Chemistry, 295(44), 14807-14825.

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