Sephadex G-25 does not have a biological target in the pharmacological sense. It is a chromatography medium used for the physical separation of molecules based on size. The "target" in a research context is the separation of biomolecules of different molecular weights. Sephadex G-25 has a separation range of approximately 1 to 5 kDa for globular proteins. It is used to separate small molecules (salts, buffer components, small peptides) from larger molecules (proteins, nucleic acids) through size exclusion.

Guidelines (This is our suggested protocol; it should be adjusted based on your unique requirements as it just offers as a guideline). 1. Swelling: It is necessary to swell the dry rubber particles initially. Stem excess deionized water and let it swell for three hours at room temperature or for one hour in a water bath at ninety degrees Celsius. (To prevent fracturing the microspheres during the swelling phase, magnetic stirring is prohibited.) 2. Column packing: (1) Based on the characteristics of the samples to be separated, prepare the elution and initial buffers (equilibrium solution and initial buffer). It is usually advised to utilize a single buffer with a low salt concentration for sample loading, equilibration, and elution in gel chromatography for desalting proteins, such as proteins. For instance, 50 mM Tris-hydrochloric acid or PBS buffer close to neutrality; if peptide separation is the goal, 0.15 M NaCl can be suitably added to prevent target-gel contact; if buffer exchange is the goal, choose the target buffer as required. (2) After draining the gel, make a homogenate with the first buffer (3:1), then degas. (3) Fix the column vertically, moisten the lower end with buffer or water, and maintain the liquid level for a while. (4) Using a glass rod as a guide, pour the homogenate into the column one time around the inner wall, letting the gel settle freely. Then, cover the column with water and let it settle for the night. (5) Attach the movable column head at the top of the column, activate the peristaltic pump, allow the buffer to flow through five times the column volume using the operating flow rate, and then add 1.5 times the operating flow rate to allow the buffer to flow through five times the column volume, adjusting the adapter head to the closest setting. Lastly, use two to three column volumes of buffer to equilibrate the column. Note: To guarantee the gel's homogeneity, air bubbles cannot be added during any step of the process. 3. Equilibration: Using the equilibration buffer, equate the column at the operating flow rate. Then, watch how the detector changes until the conductivity and pH levels don't change. 4. To load a sample, turn on the switching valve. The type of sample and chromatography medium quantity are taken into consideration while choosing the sample load. In general, the sample load doesn't go above 25% of the volume of the column bed. The samples underwent pretreatment by filtering through a 0.45 μm microporous membrane after being prepared with an equilibration solution. 5. Elution: After peaking, switch to an elution buffer to achieve consistent elution to baseline equilibrium. 6. Regeneration: Rinse with deionized water for three to five times the column volume after washing with 0.2 mol/L NaOH or non-ionic detergent for two times the column volume. 7. Storage: After using, swap out the mobile phase for 20% ethanol and keep the mixture in a 4°C refrigerator. 8. Maintaining column heights of various particle sizes at 30 cm and above for coarse particles, 20 cm and above for medium particles, 10 cm and above for fine particles, and 5 cm and above for ultra-fine particles is advised in order to achieve an improved desalination effect. The smaller the particle size, the slower the flow rate and the higher the resolution. 9. The experiment should be used to determine how much sample is added. The ideal sample load is between 10% and 20% of the column bed volume, with a maximum load of 30% not to be exceeded.

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Sephadex G-25 is not a biologically active compound but a chromatography medium used in vitro for sample preparation and purification. The resin's activity is measured by its separation efficiency, which is determined by the fractionation range (1-5 kDa for globular proteins), flow rate, and resolution. Sephadex G-25 is suitable for buffer exchange and desalting of oligonucleotides for use in sequencing, PCR, and labeling applications. It is also used to isolate peptides and to remove low molecular weight compounds from reaction mixtures.

In vivo, Sephadex G-25 is not used as a therapeutic agent. It is a laboratory reagent used exclusively for in vitro applications in biochemistry and molecular biology. The resin is not administered to animals or humans. Its applications are limited to sample preparation, protein purification, buffer exchange, and desalting in research and diagnostic laboratories.

Sephadex G-25 is a chromatography medium and is not used in standard binding or receptor assays. Its properties are characterized by its swelling capacity, particle size, fractionation range (1-5 kDa for globular proteins), and flow characteristics. The resin is typically packed into a chromatography column and equilibrated with an appropriate buffer. Samples are applied, and molecules are eluted based on size. The resin's performance is evaluated by the separation of standard proteins or by the removal of salts from a sample.

Sephadex G-25 is not used in cell-based assays as it is a chromatography medium. However, samples prepared using Sephadex G-25 (e.g., desalted proteins or nucleic acids) may be used in subsequent cellular assays. The resin itself does not interact with cells and is not added to cell cultures. Its role is in sample preparation for downstream applications including cell-based experiments, molecular biology assays, and protein analysis.

Sephadex G-25 is not used in animal experiments. It is a laboratory reagent for in vitro sample preparation. The resin is not administered to animals. Its use is limited to the preparation of samples (e.g., proteins, peptides, nucleic acids) that may be used in animal studies. The resin itself is not tested in vivo.

Pharmacokinetic properties are not applicable to Sephadex G-25 as it is not a therapeutic agent. It is a chromatography medium used for sample preparation. The resin's physical and chemical properties including particle size, swelling capacity, and chemical stability are characterized for chromatography applications. The resin is typically stored dry and swells in aqueous buffers before use. No PK data are available or relevant.

The toxicity of Sephadex G-25 has not been evaluated as it is not a therapeutic agent. The resin is considered non-toxic for laboratory use when handled appropriately. As a cross-linked dextran polymer, it is chemically inert and biocompatible. Standard laboratory safety practices should be followed when handling the resin, including the use of gloves and eye protection. The resin is for research use only and is not intended for human consumption or therapeutic use.

[1]. A rapid method of concentrating proteins in small volumes with high recovery using Sephadex G-25. Anal Biochem. 1984 May 1;138(2):451-3.

Sephadex G-25 (CAS# 9041-35-4) is a cross-linked dextran gel filtration medium used for the separation of molecules based on size. It has a fractionation range of 1 to 5 kDa for globular proteins and is suitable for desalting, buffer exchange, and separation of peptides, proteins, and oligonucleotides. Sephadex G-25 is widely used in molecular biology for buffer exchange of oligonucleotides for sequencing, PCR, and labeling applications, as well as for the isolation of peptides from complex mixtures. It is a laboratory reagent and is not approved as a therapeutic agent.

0

272.11

9041-35-4

3084046

White to off-white solid powder

410.8ºC at 760mmHg

8

9

3

18

176

5

C([C@@H]1[C@H]([C@H]([C@H]([C@@H](O1)O)O)O)O)O.C(C(CO)O)O

BCPVZFFJGCURNR-OCOFDJSDSA-N

InChI=1S/C6H12O6.C3H8O3/c7-1-2-3(8)4(9)5(10)6(11)12-2;4-1-3(6)2-5/h2-11H,1H2;3-6H,1-2H2/t2-,3-,4-,5-,6-;/m1./s1

(2R,3R,4R,5S,6R)-6-(hydroxymethyl)oxane-2,3,4,5-tetrol;propane-1,2,3-triol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)