Mini dialysis kit

Manufactured by GE Healthcare

Sourced in United States

The Mini Dialysis Kit is a lab equipment product designed for the efficient removal and exchange of small molecules in solution. It functions as a compact dialysis system, enabling the purification and concentration of samples.

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Lab products found in correlation

Ni nta agarose, by Fujifilm (2 mentions) Amicon ultra device, by Merck Group (2 mentions) Imidazole, by Fujifilm (2 mentions) Glutathione sepharose 4b bead, by Cytiva (1 mentions) 1 2 dioleoyl sn glycero 3 phosphoethanolamine, by Avanti Polar Lipids (1 mentions) N isopropylmethacrylamide nipmam, by Merck Group (1 mentions) Methanol, by GE Healthcare (1 mentions) Chloroform, by GE Healthcare (1 mentions) Hexadecyltrimethylammonium bromide ctab, by Merck Group (1 mentions) Alexa fluor 488 nhs ester, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PlusOne Mini Dialysis Kit (3 citations)

7 protocols using mini dialysis kit

Synthesis and Purification of ALN-PEG-CSA-90 Conjugate

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CSA-90 was conjugated to the carboxyl terminal of ALN-PEG-COOH using water soluble carbodiimide catalyst. Briefly, EDC.HCl (30 mg, 157 µmol) and CSA-90 (25.5 mg, 36.2 µmol) were added to the solution of ALN-PEG-COOH. The reaction mixture was then stirred for 24 h at room temperature. The resultant ALN-PEG-CSA-90 conjugate (BBA-1) was dialyzed for 24 h using a Mini Dialysis Kit (1 kDa cut-off) obtained from GE Healthcare Bio-sciences, Princeton, NJ, USA (catalogue number 80-6483-94). The goal was to remove any unconjugated CSA-90 and excess of EDC.HCl or any intermediate product formed during the reaction. The purified product was subsequently freeze-dried for use in further studies.

Kamble S., Valtchev P., Dao A., Pelras T., Rogers M.J., Savage P.B., Dehghani F, & Schindeler A. (2021). Synthesis and Characterization of Bone Binding Antibiotic-1 (BBA-1), a Novel Antimicrobial for Orthopedic Applications. Molecules, 26(6), 1541.

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Dialysis of DFCM-FM-3 for Purification

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Dialysis of DFCM-FM-3 was done using a Mini Dialysis Kit (GE Healthcare, UK) to remove small molecules (<1 kD). The dialysis tube and cap were rinsed with distilled water. DFCM-FM-3 was added to the tube. The dialysis tube was inverted into a beaker containing DPBS without antibiotics overnight at 4 °C. The tube was spun in a magnetic stirrer briefly before collecting the dialysed DFCM-FM-3, denoted as DFCM-FM-3 (dialysis).

Abdul Ghani N.‘., Razali R.A., Chowdhury S.R., Fauzi M.B., Bin Saim A., Ruszymah B.H, & Maarof M. (2022). Effect of Different Collection Times of Dermal Fibroblast Conditioned Medium (DFCM) on In Vitro Re-Epithelialisation Process. Biomedicines, 10(12), 3203.

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Recombinant Protein Expression and Purification

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Recombinant GST- or His₆-tagged proteins were expressed in and purified from E. coli. The BL21(DE3)pLysS bacterial cells were transformed with pGEX-6P- or pET30-based vectors, cultured, and then exposed to 0.5 mM isopropyl-β-d-thiogalactopyranoside for 16 h at 10 °C. The cells were then subjected to ultrasonic treatment, and the soluble fraction of cell lysates was purified. The expressed GST-tagged proteins were purified with glutathione-Sepharose 4B beads (Amersham Biosciences) and were eluted from the beads with reduced glutathione. The expressed His₆-tagged proteins were purified with Ni-NTA agarose (Wako) and were eluted with imidazole (Wako). Purified proteins were concentrated with an Amicon Ultra device (Merck Millipore, Tokyo, Japan) and dialyzed against PBS with a Mini Dialysis Kit (GE Healthcare).

Shirane M., Wada M., Morita K., Hayashi N., Kunimatsu R., Matsumoto Y., Matsuzaki F., Nakatsumi H., Ohta K., Tamura Y, & Nakayama K.I. (2020). Protrudin and PDZD8 contribute to neuronal integrity by promoting lipid extraction required for endosome maturation. Nature Communications, 11, 4576.

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Pronase E Digestion of Glycoproteins

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Twelve mg of RNase B or 30.6 mg of A. niger secretome dissolved in NH₄HCO₃ buffer (100 mM, pH 8) were reduced with 10 mM dithiothreitol at 60 °C for 30 min and then alkylated with 40 mM iodoacetamide in the dark at 37 °C for 60 min. With the addition of 1 mM CaCl₂, the chemically treated proteins were digested with pronase E from S. griseus (mass of protein substrate/mass of pronase E = 1/1) at 37 °C for 6 h and further digested with newly added fresh pronase E at 37 °C for 18 h. The concentration of the digested peptides was determined using bicinchoninic acid (BCA) assay. The digestion efficiency was examined by running a gel with the loading amount of 10 μg. The completely or nearly completely digested products were used in the downstream experiments. Portion of the pronase E digest of A. niger secretome was dialyzed with Mini Dialysis Kit (1 kDa cutoff, GE Healthcare Life Sciences, Pennsylvania) 3 times with the total dialysis time of 17 h.

Qu Y., Feng J., Deng S., Cao L., Zhang Q., Zhao R., Zhang Z., Jiang Y., Zink E.M., Baker S.E., Lipton M.S., Paša-Tolić L., Hu J.Z, & Wu S. (2014). Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection. Fungal genetics and biology : FG & B, 72, 207-215.

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Recombinant Protein Purification in E. coli

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Recombinant His₆-tagged proteins were expressed in and purified from Escherichia coli. The BL21(DE3)pLys bacterial cells were transformed with pET30-based vectors, cultured, and then exposed to 0.5 mM isopropyl-β-D-thiogalactopyranoside for 16 h at 10°C. The cells were then subjected to ultrasonic treatment, and the soluble fraction of the cell lysates was isolated. The expressed His₆-tagged proteins were purified with the use of Ni-NTA agarose (Wako) and were eluted with imidazole (Wako). The purified proteins were concentrated with an Amicon Ultra device (Merck Millipore) and dialyzed against PBS with the use of a Mini Dialysis Kit (GE Healthcare).

Morita K., Wada M., Nakatani K., Matsumoto Y., Hayashi N., Yamahata I., Mitsunari K., Mukae N., Takahashi M., Izumi Y., Bamba T, & Shirane M. (2022). PDZD8-deficient mice accumulate cholesteryl esters in the brain as a result of impaired lipophagy. iScience, 25(12), 105612.

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Bioconjugation of Peptides and Proteins to Quantum Dots

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An amount of 2 µL water-soluble QD-COOH solution (8 µM CdSe-ZnS) was added to 2.5 mL of phosphate buffer solution (0.0067 M, pH = 7.0–7.2), and then 2 µL of EDC solution (20 mg/mL N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride in PBS) was added to the QDs phosphate buffer solution. A total of 10 µL of peptide or protein solution (10 ng/uL peptides or proteins in PBS) was added to the mixture, which was placed at room temperature for 8 h to promote coupling between the peptides or proteins and QDs. After peptides or proteins binding, excess peptide molecules or proteins were removed using a mini dialysis kit (GE Healthcare, Chicago, FL, USA). Finally, 2 µL of BSA (20 mg/mL albumin from bovine serum in PBS) was added to the mixture, and the mixture was placed at room temperature for another 8 h to block the QDs. Through the above steps, a solution of QDs coupled with peptides or proteins was obtained.

Zheng Y., Song K., Cai K., Liu L., Tang D., Long W., Zhai B., Chen J., Tao Y., Zhao Y., Liang S., Huang Q., Liu Q., Zhang Q., Chen Y., Liu Y., Li H., Wang P., Lan K., Liu H, & Xu K. (2022). B-Cell-Epitope-Based Fluorescent Quantum Dot Biosensors for SARS-CoV-2 Enable Highly Sensitive COVID-19 Antibody Detection. Viruses, 14(5), 1031.

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Liposome Preparation and Labeling

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N-isopropylmethacrylamide (NIPMAM; Sigma-Aldrich), N,N′-methylenebisacrylamide (BIS; Fluka), hexadecyltrimethylammonium bromide (CTAB; Sigma-Aldrich), and 2,2′-Azobis(2-methylpropionamidine) dihydrochloride (V50; Fluka) were used as received. Styrene monomer (BASF) was purified on an Al₂O₃ column prior to use. The following were purchased from Avanti Polar Lipids: 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC, C₄₄H₈₄NO_8P), 1,2-ditetradecanoyl-sn-glycero-3-phosphocholine (DMPC, C₃₆H₇₂NO_8P), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (ammonium salt) (Liss Rhod PE, C₆₈H₁₀₉N₄O₁₄PS₂). Alexa Fluor^TM 488 NHS Ester (C₂₅H₁₅Li₂N₃O₁₃S₂) was purchased from Thermo Fisher Scientific Inc. Cholesterol (C₂₇H₄₆O), chloroform (analytical grade, ≥ 99.8%), methanol (analytical grade, ≥ 99.9%), Mini Dialysis Kit (molecular weight cutoff, 8 kDa; GE Healthcare) were purchased from Sigma-Aldrich. MilliQ water was used in all the experiments.

Liu X., Auth T., Hazra N., Ebbesen M.F., Brewer J., Gompper G., Crassous J.J, & Sparr E. (2023). Wrapping anisotropic microgel particles in lipid membranes: Effects of particle shape and membrane rigidity. Proceedings of the National Academy of Sciences of the United States of America, 120(30), e2217534120.

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