Spin columns Lectin Kits - glycoproteins - micro columns

Description

AffiSpin-Kits are designed especially for the purification of microgram amounts of glycoproteins. They are ready-to-use and thus provide easy handling. They combine AffiSpin micro columns with the corresponding optimized adsorption and elution buffers and collection tubes. The Kits are designed for operation in a centrifuge, vacuum manifold or with the provided syringes. They facilitate glycoprotein separations in a simple way for fast and reproducible results.

AffiSpin columns are packed with 0.1 mL adsorbent.
The Kit is designed for two affinity separations.

Fig: Cap and plug of an AffiSpin column are removed. AffiSpin columns are placed into an open 2 mL collection tube. A sample is dissolved in the corresponding adsorption buffer A and added to the column.

Fig: The AffiSpin column with a collection tube is placed into a centrifuge. Separation is performed by spinning the open column for 2 min at 500 U/min.

Application 308:

Binding capacity of AffiSpin ConA for the glycoprotein ovalbumin

Column:	AffiSpin ConA column (Cat. No. 051041); 0.1 mL adsorbent
Mobile Phase A:	ConA adsorption buffer
Mobile Phase B:	ConA elution buffer B2
Detection:	Bradford protein assay
Sample:	Ovalbumin; 400 µg dissolved in mobile phase A

The table below shows the results of the lectin affinity separation of the glycoprotein ovalbumin on AffiSpin ConA. 400 µg ovalbumin were added to the AffiSpin ConA column. Flow through, elution and washing steps were performed as described in the protocol for using AffiSpin lectin columns and the fractions were collected and pooled. 187 µg of ovalbumin were bound by the column and could be specifically eluted and recovered.

	Total protein applied	Flow through	Elution and washing	Total recovery
Amount	400 µg	161 µg	187 µg	348 µg
Recovery		40%	47%	87%

Application 311:

Separation of the glycoproteins ovalbumin and ovomucoid on AffiSpin ConA

Column:	AffiSpin ConA (Cat. No. 051041)
Sample:	Ovalbumin; 200 µg dissolved in A Ovomucoid; 200 µg dissolved in A
Mobile Phase A:	ConA adsorption buffer
Mobile Phase B:	ConA elution buffer B2
Detection:	SDS-PAGE

The picture stated below shows the results of the separation of a mixture of two glycoproteins with AffiSpin ConA. 200 µg ovalbumin and 200 µg ovomucoid dissolved in adsorption buffer were mixed and added to the used AffiSpin ConA column. The column was treated as described in the protocol for using AffiSpin lectin columns. Afterwards the samples were analyzed with SDS-PAGE. The gel was stained with Coomassie stain.

M
3
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6
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8

Legend

Molecular weight marker
Ovalbumin
Ovomucoid
Mixture of ovalbumin and ovomucoid applied to the spin column
Flow through
Elution fraction
Washing step

AffiSpin® Kits

AffiSpin^® Kits