Protein L Agarose Resin

Protein L is an immunoglobulin-binding protein that was isolated from the bacteria Peptostreptococcus magnus and provides a convenient way to separate immunoglobulins from a variety of sources. Recombinant Protein L contains four immunoglobulin binding domains of the native protein, and recombinant Protein L resins can be used for the purification of IgG, IgM, IgA and IgD containing kappa light chains from various species without interfering with the antigen binding site. Besides antibodies, Protein L is also suitable for binding of a wide range of antibody fragments such as Fabs, single-chain variable fragments (scFv), and domain antibodies (Dabs). Protein L is immobilized by means of covalent binding that avoids protein loss and allows for column re-use.

Bead Geometry & Size	Spherical ,Standard: ~50-150 µm
Crosslinked	Yes
Agarose %	4%
Coupling Method	Covalent binding by reductive amination
Static Binding Capacity	~10 mg human IgG/ ml resin
Antimicrobial Agent	20% Ethanol

Target	Protein L Agarose Resin
Storage	Store at 2-8°C.
Specificity	Affinity of Protein L for Ig types: Species Ig Subtype Affinity for Protein L Species Ig Subtype Affinity for Protein L Human Total IgG +++ Rabbit Total IgG + Human IgG1 ++++ Chicken IgY + Human IgG2 ++++ Cow Total IgG - Human IgG3 +++ Cow IgG1 - Human IgG4 ++++ Cow IgG2 - Human IgA +++ Goat Total IgG - Human IgA1 +++ Goat IgG1 - Human IgA2 +++ Goat IgG2 - Human IgD +++ Sheep Total IgG - Human IgE ++++ Sheep IgG1 - Mouse Total IgG +++ Sheep IgG2 - Mouse IgG1 +++ Hamster - +++ Mouse IgG2a +++ Horse Total IgG Not Determined Mouse IgG2b +++ Cat Total IgG Not Determined Mouse IgG3 +++ Dog - Not Determined Mouse IgM +++ Pig - +++ Rat Total IgG +++ Guinea pig IgG1 Not Determined Rat IgG1 +++ Guinea pig IgG2 Not Determined Rat IgG2a +++ Koala - Not Determined Rat IgG2b +++ Llama - Not Determined Rat IgG2c +++ Monkey (Rhesus) - Not Determined Rat IgG3 Not Determined
Availability	Shipped within 5-10 working days.
Note	THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use	Suggested Protocol Gravity Purification - It is recommended to use empty plastic gravity columns to carry out this procedure. Remove Preservatives: Determine the quantity of Protein L Agarose needed for the intended purification. Gently shake the bottle of Protein L Agarose to achieve a homogenous suspension. Immediately pipette the required volume of suspension to an empty column. Remove the upper cap of the column, and then the lower cap, to allow the elimination of preservatives by gravity flow. Equilibrate Pre-packed Columns: Equilibrate the column with 5 bed volumes of binding buffer (0.1 M phosphate, 0.15 M NaCl, pH 7.2). Add the binding buffer to the upper part of the column. Ensure that there is no air trapped. Replace the cap on the column and mix by inverting the column, then discard the supernatant. Repeat this step two more times for a total of three times. Add Sample: Dilute the sample with binding buffer. For optimal binding, it is recommended to determine and maintain the optimal ionic strength and pH. For plasma samples only, after dilution, centrifuge at 10,000 × g for 20 minutes, then take the supernatant. Add the sample through the top of the column. Allow the sample and resin to remain in contact for at least 1 hour. In some cases, a slight increase in contact time may facilitate binding. Binding capacity can be affected by several factors such as sample concentration. Replace the cap on the column and mix by inverting the column, then remove the lower cap of the column and discard the supernatant. Wash Pre-packed Columns: Close the column outlet with the cap. Add 10 bed volumes of binding buffer through the top of the column. Close the column inlet with the cap, then mix by inverting the column. Remove the lower cap of the column and discard the supernatant. Repeat this step two more times for a total of three times. The OD280 of the supernatant should be the same as the OD280 of pure binding buffer when the washing process is complete. Elute Pure Immunoglobulins: Close the column outlet with the cap. Add 1 bed volume of elution buffer (0.1 M glycine, pH 2.0; citric acid buffer, pH 2.8; or other suitable reduced pH buffer) to the column. Close the column inlet with the cap and mix thoroughly by inverting the column for at least 10 min at room temperature. Sediment the gel, remove the end cap and collect the eluate in a new tube and store on ice. Repeat this step twice for a total of three times, and pool the collected eluates. It is recommended to add 0.15 ml of pH 9.0 buffer (e.g 1 M Tris) per ml of purified immunoglobulin to neutralize the eluted fractions. Store the combined eluate between 2-8 °C in a suitable bacteriostat (e.g. 20% ethanol). Do not freeze the eluate. Batch Purification Remove Preservatives: Determine the quantity of Protein L Agarose needed for the intended purification. Gently shake the bottle of Protein L Agarose to achieve a homogenous suspension. Immediately pipette 2 ml of Protein L Agarose suspension per ml of gel volume required to an empty tube. Sediment the gel by centrifugation at 500 × g for 5 minutes, then carefully decant the supernatant and discard it. Equilibrate Resin: Equilibrate the gel with 10 bed volumes of binding buffer (0.1 M phosphate, 0.15 M NaCl, pH 7.2). Sediment the gel by centrifugation at 500 × g for 5 minutes, then carefully decant the supernatant and discard it. Add Sample: Add the sample to the resin and mix the suspension gently for 1 hour at room temperature. In some cases, a slight increase in contact time may facilitate binding. Centrifuge the suspension at 500 × g for 5 minutes to sediment the resin, then carefully decant the supernatant and discard it. Wash Resin: Add 10 bed volumes of binding buffer through the top of the column. Mix by inverting the suspension. Centrifuge the suspension at 500 × g for 5 minutes to sediment the resin, then carefully decant the supernatant and discard it. Repeat this step two more times for a total of three times. The OD280 of the supernatant should be the same as the OD280 of pure binding buffer when the washing process is complete. Elute Pure Immunoglobulins: Add 1 bed volume of elution buffer (0.1 M glycine, pH 2.0; citric acid buffer, pH 2.8; or other suitable reduced pH buffer) to the gel. Mix thoroughly for 10 min at room temperature. Sediment the gel by centrifugation at 500 × g for 5 minutes, then carefully decant the supernatant into a new tube and store on ice. Repeat this step twice for a total of three times, and pool the collected eluates. It is recommended to add 0.15 ml of pH 9.0 buffer (e.g 1 M Tris) per ml of purified immunoglobulin to neutralize the eluted fractions. Store the combined eluate between 2-8 °C in a suitable bacteriostat (e.g. 20% ethanol). Do not freeze the eluate.

Research Articles on Protein L Agarose Resin

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