Beh125 sec protein standard mix

Manufactured by Waters Corporation

Sourced in United States

The BEH125 SEC Protein Standard Mix is a laboratory equipment product designed for size exclusion chromatography (SEC) applications. It provides a set of well-characterized protein standards to assist in the calibration and performance evaluation of SEC columns and instrumentation. The product includes a mixture of proteins with a range of molecular weights, allowing users to establish molecular weight-based separation profiles and verify the functionality of their SEC system.

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Lab products found in correlation

Acquity 1 class uplc instrument, by Waters Corporation (2 mentions) Acquity beh125 sec column, by Waters Corporation (1 mentions) Empower 3, by Waters Corporation (1 mentions) Nexera xr, by Shimadzu (1 mentions)

3 protocols using beh125 sec protein standard mix

Size-Exclusion UHPLC Analysis of Allergen

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Example 8

Size-exclusion ultra-high-performance liquid chromatography (SE-UHPLC) analysis of rEqu c 1 wt and mutants was performed using Acquity BEH125 SEC column with dimensions of 4.6×150 mm, a pore size of 125 Å and a particle size of 1.7 μm (Waters) coupled with Acquity I-Class UPLC instrument (Waters). The column was equilibrated to running conditions with PBS (12 mM Na₂HPO₄, 3 mM NaH₂PO₄, 150 mM NaCl pH 7.3) as a mobile phase at flow rate 0.3 ml/min until the baseline was stable. Sample amounts of 2 μl from a 40 μM protein solution were injected. The chromatographic separation was carried out under isocratic flow with overall run time of 12 minutes and detection at 214 nm wavelength. For gel filtration standard, the BEH125 SEC Protein Standard Mix (Waters) was used. SE-UHPLC results show that the hypoallergen mutants (Triple 2, 3, and 4) exist mainly as monomers whereas wild type exists mainly as a dimer at the concentration of 40 μM (FIG. 7).

US11679152B2. Recombinant hypoallergenic Equ c 1 polypeptides for use in the immunotherapy of horse allergy (2023-06-20). Desentum Oy [FI]. Inventors: Juha Rouvinen [FI], Kristiina Takkinen [FI], Marja-Leena Laukkanen [FI], Merja Niemi [FI], Janne Jänis [FI], Jaana Haka [FI].

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SE-UHPLC Analysis of rEqu c 1 Variants

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Size-exclusion ultra-high-performance liquid chromatography (SE-UHPLC) analysis of rEqu c 1 wt and Triple variants were performed using Acquity BEH125 SEC column with dimensions of 4.6 × 150 mm, a pore size of 125 Å and a particle size of 1.7 µm (Waters) coupled with Acquity I-Class UPLC instrument (Waters) and controlled by Empower 3 software (Waters). The column was equilibrated to running conditions with PBS (12 mM Na₂HPO₄, 3 mM NaH₂PO₄, 150 mM NaCl, pH 7.3) as a mobile phase at a flow rate 0.3 ml/min until the baseline was stable. Samples of 2 µl of a 40 µM protein solution were injected. The chromatographic separation was carried out under isocratic flow with overall run time of 12 minutes and detection at 214 nm wavelength. The BEH125 SEC Protein Standard Mix (Waters) was used as a gel filtration standard.

Haka J., Niemi M.H., Mattila P., Jänis J., Takkinen K, & Rouvinen J. (2019). Development of hypoallergenic variants of the major horse allergen Equ c 1 for immunotherapy by rational structure based engineering. Scientific Reports, 9, 20148.

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Size-Exclusion UHPLC Analysis of Powdered Samples

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50 mg of each powdered CM sample was dissolved in 1 ml of mmol/L phosphate buffer pH 6.8 which was also used as mobile phase. Samples were vortexed (15 s) and left 1 h with gentle rocking at room temperature. After centrifugation (12,300×g, 10 min) supernatants were defatted by dichloromethane extraction (1:1 v/v). Prior to SE-UHPLC analysis protein concentration in the samples was determined by the Bradford assay and samples were diluted to 0.35 mg/ml. SE-UHPLC analyses were performed using ACQUITY UPLC Protein BEH SEC 125 Å column (4.6 × 150 mm I.D., 1.7 μm, Waters, Milford, MA, USA) connected to ultra-high-performance liquid chromatography (UHPLC) workstation Nexera XR (Shimadzu Corporation, Kyoto, Japan). Mobile phase flow was 0.3 ml/min and column temperature was set at 30 ̊C. UV detection was done at 220 nm wavelength. The BEH125 SEC Protein Standard Mix (Waters, Milford, MA, USA), four component protein mixture (Thyroglobulin, Ovalbumin, Ribonuclease A and Uracil) were used for column calibration. Calibration curve for column was generated after three consecutive runs. The method run time was min. Injection volumes were 4 μl for BEH SEC 125s Å column. The chromatographic control, data acquisition and analysis were performed using LabSolutions CS Analysis Data System (Shimadzu Corporation, Kyoto, Japan) version 5.73.

Khulal U., Ghnimi S., Stevanović N., Rajković A., & Veličković T.Ć. (2021). Aggregability and digestibility study of fruit juice fortified camel milk powder proteins. LWT, 152, 112250-112250.

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