Sephadex g 25

Manufactured by GE Healthcare

Sourced in United States, Sweden, United Kingdom, Germany, Japan

Sephadex® G-25 is a gel filtration medium used in size exclusion chromatography. It is a cross-linked dextran-based material that separates molecules based on their size and molecular weight. Sephadex® G-25 is commonly used for desalting, buffer exchange, and sample clean-up applications.

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Lab products found in correlation

Pd 10 desalting column, by GE Healthcare (7 mentions) Cyclone storage phosphor system, by PerkinElmer (3 mentions) Alexa fluor 488 c5 maleimide, by Thermo Fisher Scientific (3 mentions) Maximadry, by Thermo Fisher Scientific (3 mentions) Vcx130, by Sonics (3 mentions) Pgex 6p1 vector, by Thermo Fisher Scientific (3 mentions) Prescission protease, by GE Healthcare (3 mentions) Emulsiflex c5 homogenizer, by Avestin (3 mentions) Amicon ultra centrifugal filter, by Merck Group (3 mentions) Pd 10 column, by GE Healthcare (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Alexa fluor 647, by Thermo Fisher Scientific (2 mentions) Iodine 125 125i, by PerkinElmer (2 mentions) Aca34, by Merck Group (2 mentions) Hen egg ovalbumin, by GE Healthcare (2 mentions) Iodine 125, by PerkinElmer (2 mentions) Imidazole, by Sangon (2 mentions) Sodium acetate, by Sangon (2 mentions) Magnesium chloride, by Sangon (2 mentions) Ethanol, by Sangon (2 mentions)

Close spelling variants of this product

Sephadex G-25 medium (35 citations) Sephadex G-25 resin (33 citations) Sephadex G-25 PD-10 column (26 citations) Sephadex G-25 desalting column (16 citations) PD-10 Sephadex G-25 (6 citations) Sephadex G-25 gel (6 citations) NAP-10 Sephadex G-25 column (6 citations) Sephadex G-25 medium columns (5 citations) Sephadex G-25 prepacked PD10 columns (4 citations) G-25 Sephadex columns (4 citations)

231 protocols using sephadex g 25

Preparation and Characterization of Anti-CD123 ADCs

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Example 323

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Under argon, a solution of 0.029 mg of TCEP in 50 μl of PBS buffer was added to 5 mg of anti-CD123 AK_1A(TPP-5969) in 450 μl of PBS (c=11.11 mg/ml). The mixture was stirred at RT for 30 min. 0.328 mg (0.00023 mmol) of intermediate F323, dissolved in 50 μl of DMSO, was then added. After a further 90 min of stirring at RT, the mixture was diluted with 1950 μl of PBS buffer and then purified on PD-10 columns (Sephadex® G-25, GE Healthcare). The mixture was then concentrated by ultracentrifugation and rediluted with PBS buffer. The ADC batch obtained was characterized as follows:

Protein concentration: 2.03 mg/ml

Drug/mAb ratio: 2.9

Under argon, a solution of 0.029 mg of TCEP in 50 μl of PBS buffer was added to 5 mg of anti-CD123 AK_1C(TPP-6013) in 450 μl of PBS (c=11.11 mg/ml). The mixture was stirred at RT for 30 min. 0.328 mg (0.00023 mmol) of intermediate F323, dissolved in 50 μl of DMSO, was then added. After a further 90 min of stirring at RT, the mixture was diluted with 1950 μl of PBS buffer and then purified on PD-10 columns (Sephadex® G-25, GE Healthcare). The mixture was then concentrated by ultracentrifugation and rediluted with PBS buffer. The ADC batch obtained was characterized as follows:

Protein concentration: 2.06 mg/ml

Drug/mAb ratio: 4.2

US10744205B2. Antibody drug conjugates of kinesin spindel protein (KSP) inhibitors with anti-CD123-antibodies (2020-08-18). Bayer Pharma Aktiengesellschaft [DE]. Inventors: Hans-Georg Lerchen [DE], Anne-Sophie Rebstock [FR], Yolanda Cancho Grande [DE], Sven Wittrock [DE], Beatrix Stelte-Ludwig [DE], Christoph Mahlert [DE], Simone Greven [DE], Stephan Märsch [DE], Dennis Kirchhoff [DE].

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Antibody-Drug Conjugate Synthesis Using TCEP and R28

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Example 2811

Under argon, a solution of 0.17 mg of TCEP in 0.25 ml of PBS buffer (pH 7.2) was added to 30 mg of the anti-TWEAKR antibody TPP-7007 in 2.5 ml of PBS (c=12 mg/ml). The mixture was stirred at RT for 30 min, and then 1.57 mg (0.0014 mmol) of Intermediate R28 dissolved in 250 μl of DMSO was added. After stirring at RT for a further 90 min, the mixture was diluted to 5 ml with PBS buffer which had been adjusted to pH 8 beforehand, divided and then each portion was passed through a PD 10 column (Sephadex® G-25, GE Healthcare) equilibrated with PBS buffer pH 8, and eluted with PBS buffer pH 8. The eluates were combined, diluted to 7.5 ml with PBS buffer pH 8 and stirred under argon at RT overnight. This solution was then applied to PD 10 columns (Sephadex® G-25, GE Healthcare) which had been equilibrated with PBS buffer pH 7.2 and was eluted with PBS buffer pH 7.2. The eluate was then concentrated by ultracentrifugation and rediluted with PBS buffer (pH 7.2) and reconcentrated and sterile-filtered again. The ADC batch obtained was characterized as follows:

Protein concentration: 8.2 mg/ml

Drug/mAb ratio: 3.1

US11685714B2. Prodrugs of cytotoxic active agents having enzymatically cleavable groups (2023-06-27). BAYER PHARMA AKTIENGESELLSCHAFT [DE]. Inventors: Hans-Georg Lerchen [DE], Anne-Sophie Rebstock [FR], Leo Marx [CH], Sarah Anna Liesa Johannes [DE], Beatrix Stelte-Ludwig [DE], Lisa Dietz [DE], Carsten Terjung [DE], Christoph Mahlert [DE], Simone Greven [DE], Anette Sommer [DE], Sandra Berndt [DE].

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Quantitative [3H]SA Binding Assay

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[³H]SA-binding assays were performed using size exclusion chromatography as described previously (Chen and Klessig, 1991 (link)). Briefly, pre-equilibrate sephadex™ G-25 (GE healthcare) with PBS buffer containing 0.1% Tween-20 overnight at 4° C. Size-exclusion column was prepared using 1 ml syringe with glass wool fiber as filter and packed with overnight equilibrated sephadex™ G-25; excess buffer was removed by centrifugation. Binding of [³H]SA with His₆-MBP-tagged NPR1, His₆-MBP-tagged FER1, MBP and no protein control was performed in PBS buffer with 100 μl reaction volume in the absence or presence of excess unlabeled SA (10,000-fold). The reaction mix was incubated on ice for 1 h, and then loaded on the column and centrifuged. The flow through was collected and dissolved in scintillation liquid and radioactivity was measured by a scintillation counter (LS6500; Beckman Coulter, Pasadena, CA). The Kd value was determined by non-linear fitting model of Michaelis-Menten equation with [³H]SA concentration from 5 to 640 nM using OriginPro 7.5 software (OriginLab, Northampton, MA).

Manohar M., Tian M., Moreau M., Park S.W., Choi H.W., Fei Z., Friso G., Asif M., Manosalva P., von Dahl C.C., Shi K., Ma S., Dinesh-Kumar S.P., O'Doherty I., Schroeder F.C., van Wijk K.J, & Klessig D.F. (2015). Identification of multiple salicylic acid-binding proteins using two high throughput screens. Frontiers in Plant Science, 5, 777.

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Doxorubicin-Loaded Liposome Preparation

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Phospholipid, Chol and DSPE-PEG_2K at the indicated molar ratio in Fig. 5a were completely dissolved by ethanol in a 100 mL round bottom glass flask. The thin lipid film was generated through evaporating the organic solvent via a rotatory evaporator (RV 10 digital, IKA®) under ultra-high vacuum (MaximaDry, Fisherbrand) for half an hour. (NH₄)₂SO₄ buffer (125 mM) was added into the flask to hydrate the lipid film under 60 °C and rotated for half an hour. The suspension solution was transferred to a tube and sonicated via a probe under 4 °C through a pulse 3/2 s on/off at a power output of 60 W (VCX130, Sonics & Materials Inc). To remove the unencapsulated (NH₄)₂SO₄, the primary liposomal solution was gone through a PD-10 column (Sephadex G-25, GE Healthcare) via PBS buffer as eluent. Afterwards, the (NH₄)₂SO₄ laden liposomes were incubated with 6 mg/mL DOX by the ratio of DOX/total lipid = 0.1/1 (w/w) under 60 °C for 1 h. The liposomal solutions were transferred to an ice bath and cooled to ~4 °C for half an hour. To get rid of the unloaded DOX, the liposomes were passed through a PD-10 column (Sephadex G-25, GE Healthcare) via PBS buffer as eluent. DLS and HPLC were utilized to measure the size, zeta potential, PDI, and drug content of the liposome samples, respectively. The DLC [Eq. (4)] and DLE [equation (5)] of DOX in the liposomes were calculated using the formulas as above:

Wang Z., Li W., Jiang Y., Park J., Gonzalez K.M., Wu X., Zhang Q.Y, & Lu J. (2024). Cholesterol-modified sphingomyelin chimeric lipid bilayer for improved therapeutic delivery. Nature Communications, 15, 2073.

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Antioxidant Peptides from Perilla Seeds

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Perilla seeds were purchased from a local seeds market in Fuzhou, China. 1,1-diphenyl-2-picryl-hydrazil (DPPH) and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammoniumsalt (ABTS) were obtained from Sigma-Aldrich Co. (Pasadena, TX, USA). Acetonitrile, formic acid and trifluoroacetic acid (TFA) were of chromatographic grade and purchased from Sinopharm Chemical Reagent Co., Ltd (Fuzhou, China). Sephadex G-25 was purchased from GE Healthcare (Gothenburg, Sweden). RPMI-1640, fetal bovine serum (FBS), penicillin and streptomycin were purchased from Xiamen Lulong Biological Technology Development Co., Ltd (Xiamen, China). Other chemicals and solvents of analytical grade were obtained from different commercial sources. The identified peptides were synthesized by GL Biochem Ltd. (Shanghai, China), with purity higher than 98% by HPLC analysis.

Yang J., Hu L., Cai T., Chen Q., Ma Q., Yang J., Meng C, & Hong J. (2018). Purification and identification of two novel antioxidant peptides from perilla (Perilla frutescens L. Britton) seed protein hydrolysates. PLoS ONE, 13(7), e0200021.

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Synthesis and Characterization of Fluorescent Polypeptides

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Dibenzocyclooctyne-Cy5 dye (Cy5-DBCO) was conjugated to azide-PLys(Gln) and azide-PLys(α-Glu) by copper-free click chemistry to afford PLys(Gln) and PLys(α-Glu) as shown in Supplementary Fig. S4, S5. For the synthesis of PLys(Gln)-100, azide-PLys(Gln)-100 (46 mg, 0.0015 mmol) and Cy5-DBCO (0.5 mg, 0.00050 mmol) was dissolved in 10 mM NaHCO₃ aqueous solution (2 mL, pH 7.4) and stirred overnight at room temperature. The reaction mixture was purified by dialysis against deionized water (MWCO: 6–8,000). After freeze-drying, the crude product was dissolved in 1 M NaCl aqueous solution and further purified by PD-10 column (Sephadex G-25, GE Healthcare Ltd., UK), followed by dialysis against deionized water (MWCO: 6–8,000) and lyophilisation. The PLys(Gln)-100 was obtained as a blue powder (32 mg, yield = 70%). PLys(Gln)-n (n = 50, 30) or PLys(α-Glu)-n (n = 100, 50, 30) were synthesized in the same procedure. The final products were characterized by size exclusion chromatography [column: Superdex 200 increase 10/300GL (GE Healthcare Ltd); eluent: 10 mM phosphate buffer (pH 7.4) containing 150 mM NaCl; flow rate: 0.6 ml/min; temperature: room temperature; detector: fluorescence (ex/em = 620 nm/670 nm)].

Yamada N., Honda Y., Takemoto H., Nomoto T., Matsui M., Tomoda K., Konno M., Ishii H., Mori M, & Nishiyama N. (2017). Engineering Tumour Cell-Binding Synthetic Polymers with Sensing Dense Transporters Associated with Aberrant Glutamine Metabolism. Scientific Reports, 7, 6077.

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Anti-TWEAKR AK1A ADC Synthesis

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Example 270

[Figure (not displayed)]

Under argon, a solution of 0.029 mg of TCEP in 50 μl of PBS buffer was added to 5 mg of anti-TWEAKR AK_1Ain 345 μl of PBS (c=14.5 mg/ml). The reaction was diluted with 2005 μl of PBS buffer which had been adjusted to pH 8 beforehand and stirred at RT for 1 h. 0.188 mg (0.00023 mmol) of Intermediate F270 dissolved in 100 μl of DMSO were then added. After a further 90 min of stirring at RT, the reaction was applied to PD 10 columns (Sephadex® G-25, GE Healthcare) which had been equilibrated with PBS buffer pH 8 and was eluted with PBS buffer pH 8. The eluate was stirred under argon at RT overnight and then concentrated by ultracentrifugation and rediluted with PBS buffer (pH 7.2). Under these conditions, some of the ADCs may also be present in the ring-closed form. The ADC batch obtained was characterized as follows:

Protein concentration: 1.49 mg/ml

Drug/mAb ratio: 2.6

US10485880B2. Antibody-drug conjugates (ADCs) of KSP inhibitors with aglycosylated anti-TWEAKR antibodies (2019-11-26). Bayer Pharma Aktiengesellschaft [DE]. Inventors: Hans-Georg Lerchen [DE], Sven Wittrock [DE], Yolanda Cancho Grande [DE], Beatrix Stelte-Ludwig [DE], Anette Sommer [DE], Sandra Berndt [DE], Christoph Mahlert [DE], Anne-Sophie Rebstock [FR], Carsten Terjung [DE], Simone Greven [DE].

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TCEP-mediated Conjugation of Intermediate F312 to Anti-TWEAKR AK1A Antibody

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Example 312

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Under argon, a solution of 0.029 mg of TCEP in 50 μl of PBS buffer was added to 5 mg of anti-TWEAKR AK_1Ain 500 μl of PBS (c=10 mg/ml). The reaction was stirred at RT for 30 min, and 0.26 mg (0.00023 mmol) of Intermediate F312, dissolved in 50 μl of DMSO, were then added. After a further 90 min of stirring at RT, the reaction was diluted with 1900 μl of PBS buffer which had been adjusted to pH 8 beforehand.

This solution was then applied to PD 10 columns (Sephadex® G-25, GE Healthcare) which had been equilibrated with PBS buffer pH 8 and was eluted with PBS buffer pH 8. The eluate was stirred under argon at RT overnight and then concentrated by ultracentrifugation and rediluted with PBS buffer (pH 7.2). Under these conditions, some of the ADC may also be present in the ring-closed form. The ADC batch obtained was characterized as follows:

Protein concentration: 1.65 mg/ml

Drug/mAb ratio: 2.0

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TWEAKR Antibody-Drug Conjugate Synthesis

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Example 279

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Under argon, a solution of 0.029 mg of TCEP in 50 μl of PBS buffer was added to 5 mg of anti-TWEAKR AK_1Ain 239 μl of PBS (c=20.9 mg/ml). The reaction was diluted with 2111 μl of PBS buffer which had been adjusted to pH 8 beforehand and was stirred at RT for 1 h. 0.242 mg (0.00023 mmol) of Intermediate F279, dissolved in 100 μl of DMSO, was then added. After a further 90 min of stirring at RT, the reaction was applied to PD 10 columns (Sephadex® G-25, GE Healthcare) which had been equilibrated with PBS buffer pH 8 and was eluted with PBS buffer pH 8. The eluate was stirred under argon at RT overnight and then concentrated by ultracentrifugation and rediluted with PBS buffer (pH 7.2). Under these conditions, some of the ADC may also be present in the ring-closed form. The ADC batch obtained was characterized as follows:

Protein concentration: 2.19 mg/ml

Drug/mAb ratio: 2.8

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Antibody-Drug Conjugate Synthesis

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Example 277

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Under argon, a solution of 0.029 mg of TCEP in 50 μl of PBS buffer was added to 5 mg of anti-TWEAKR AK_1Ain 269 μl of PBS (c=20.9 mg/ml). The reaction was diluted with 2081 μl of PBS buffer and stirred at RT for 1 h. 0.205 mg (0.00023 mmol) of Intermediate F277, dissolved in 100 μl of DMSO, was then added. After a further 20 h of stirring at RT, the reaction was passed over PD 10 columns (Sephadex® G-25, GE Healthcare) which had been equilibrated with PBS buffer and was eluted with PBS buffer. The eluate was then concentrated by ultracentrifugation and rediluted with PBS buffer. The ADC batch obtained was characterized as follows:

Protein concentration: 1.88 mg/ml

Drug/mAb ratio: 3.3

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