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ReadiUse™ Disposable PD-10 Desalting Column

ReadiUse™ Disposable PD-10 Desalting Column packed with Sephadex G-25 resin.
ReadiUse™ Disposable PD-10 Desalting Column packed with Sephadex G-25 resin.
Ordering information
Price ()
Catalog Number60504
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Physical properties
Molecular weightN/A
SolventWater
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageRefrigerated (2-8 °C)
UNSPSC12171501
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OverviewpdfProtocol


Molecular weight
N/A
ReadiUse™ Disposable PD-10 Desalting Columns contain Sephadex G-25 resin for rapid buffer exchange, desalting, and removal of small contaminants (e.g., salts, dyes, radioactive labels) from samples using gravity flow or centrifugation. It is an inexpensive and convenient alternative to lengthy and tedious dialysis procedures. The desalting and buffer exchange is run by gravity flow (1.0 to 2.5 mL samples) or centrifugation (1.75 to 2.5 mL samples). The salt removal rate is typically >98% salt with gravity and >90% with centrifugation. The sample recovery is typically in the range of 70% to >95%.

Example protocol


SAMPLE EXPERIMENTAL PROTOCOL

Spin Protocol
  1. PD-10 Desalting column preparation

    • Cut off the tip at the notch and place the column on a rack. Remove the cap to allow the excess packing buffer to drain by gravity until reaching the top of the gel bed. If the column does not begin to flow, push the cap back into the column and remove it again to start the flow. Discard the flow-through.
    • Put the reservoir to the top of the column as shown in Figure 1.
  2. Column equilibration

    • Fill the reservoir with the equilibrium buffer of your choice (for example, PBS). Let the buffer drain out by gravity.
    • Repeat 2 times and discard the flow-through.
  3. Sample application

    • Remove the buffer reservoir and place the column in a 50 mL centrifuge tube with a spin adapter (Figure 2).
    • Centrifuge for 5 min in a swinging bucket centrifuge at 1,000x g to remove the reaction buffer. Discard the flow-through.
    • Place the column in a clean 50 mL centrifuge tube with a spin adapter.
    • Carefully apply the sample (1000 μL) directly to the top center of the column. Sample volume may need to be carefully adjusted to reach the best performance.
  4. Elution

    • After loading the sample, add 100 µL equilibrium buffer slowly in the middle of the packed bed and centrifuge the column for 10 min at 1,000 x g.
    • Collect the eluate.

Note. The ReadiUse™ PD-10 Column can fit into a 50 mL centrifuge tube with a PD-10 spin adapter for sample collection during centrifugation.

Swinging bucket centrifuges capable of generating a minimum force of 1,000g are suitable for ReadiUse™ PD-10 column use. The gravitational force created at a particular revolution speed is a function of the radius of the centrifuge rotor. Consult the swinging bucket centrifuge instruction manual for the information about conversion from revolutions per minute (RPM) to centrifugal or g-force. Alternatively, use the equation below to calculate the speed in RPM required to reach the gravitational force of 1,000 x g.

CF(g) = (1.12 x 10-5) x(RPM)2 x r

  • RCF = the relative centrifugal force
  • RPM = the speed of the rotor
  • r = the radius in centimeters measured from the center of the rotor to the middle of the PD-10 column
Gravity Protocol

The liquid passes through the column by gravity force. There is a slightly higher recovery and desalting capacity using the gravity protocol than the spin protocol, but the applied sample is diluted.

If the sample has color, you can choose the Gravity Protocol. If the sample is clear, the Spin Protocol is recommended.

  1. PD-10 Desalting column preparation

    • Cut off the tip at the notch and place the column on a rack. Remove the cap to allow the excess packing buffer to drain by gravity until reaching the top of the gel bed. If the column does not begin to flow, push the cap back into the column and remove it again to start the flow. Discard the flow-through.
  2. Column equilibration

    • Fill up the column with the equilibration buffer and allow the equilibration buffer to enter the packed bed completely.
    • Repeat several times. ~25 mL equilibration buffer should be used to equilibrate the column.
    • Discard the flow-through.
  3. Sample application

    • Add 1.0 mL of sample to the column.
    • Wait until it enters the packed bed completely.
  4. Elution (volumes may need to be adjusted based on different samples)

    • Add 1.0 mL equilibration buffer at a time. After it enters the packed bed completely, add another 1.0mL.
    • Discard the first 2.0 mL flow-through solution and start to collect elution from 3.0 mL.
    • Collect ~ 2.0-3.0 mL eluate. (Note: 0.5~1 mL/fraction)

References


View all 5 references: Citation Explorer
Fully Automated 89Zr Labeling and Purification of Antibodies.
Authors: Poot, Alex J and Adamzek, Kevin W A and Windhorst, Albert D and Vosjan, Maria J W D and Kropf, Saskia and Wester, Hans-Jurgen and van Dongen, Guus A M S and Vugts, Danielle J
Journal: Journal of nuclear medicine : official publication, Society of Nuclear Medicine (2019): 691-695
Recombinant production of A1S_0222 from Acinetobacter baumannii ATCC 17978 and confirmation of its DNA-(adenine N6)-methyltransferase activity.
Authors: Blaschke, Ulrike and Suwono, Beneditta and Zafari, Sachli and Ebersberger, Ingo and Skiebe, Evelyn and Jeffries, Cy M and Svergun, Dmitri I and Wilharm, Gottfried
Journal: Protein expression and purification (2018): 78-85
Synthesis and characterization of (18)F-labeled active site inhibited factor VII (ASIS).
Authors: Erlandsson, Maria and Nielsen, Carsten H and Jeppesen, Troels E and Kristensen, Jesper B and Petersen, Lars C and Madsen, Jacob and Kjaer, Andreas
Journal: Journal of labelled compounds & radiopharmaceuticals (2015): 196-201
In vitro and in vivo evaluation of direct rhenium-188-labeled anti-CD52 monoclonal antibody alemtuzumab for radioimmunotherapy of B-cell chronic lymphocytic leukemia.
Authors: De Decker, Mario and Bacher, Klaus and Thierens, Hubert and Slegers, Guido and Dierckx, Rudi A and De Vos, Filip
Journal: Nuclear medicine and biology (2008): 599-604
Different techniques for urinary protein analysis of normal and lung cancer patients.
Authors: Tantipaiboonwong, Payungsak and Sinchaikul, Supachok and Sriyam, Supawadee and Phutrakul, Suree and Chen, Shui-Tein
Journal: Proteomics (2005): 1140-9