Warning: The amount of imidazole required in the elution buffer for efficient release of the target protein from the resin depends on various parameters: the length of the His-tag, the accessibility of the His-tag. Due to the specific characteristics of cOmplete His-Tag Purification Resin, a protein can already be eluted with approximately 25 mM imidazole. Warning: Protein peaks can be expected between 25 to 45 mM imidazole.
Warning: Since the binding specificity of the resin is high, the kinetics of adhesion of the protein to the resin is slower than other available resins. Warning: To prevent blockage of the resin, remove insoluble material prior to loading the column.
USE HIGH S RESIN FULL
Buffers recommended in this document are well established examples and can be adapted to achieve optimal conditions for a specific target protein.ĬOmplete His-Tag Purification Resin offers full flexibility in selecting the optimal buffer conditions without compromises. Purification of native protein should be performed using optimal buffer conditions for the target protein.
Let the excess buffer drain through the column by gravity flow.Transfer the appropriate amount of resin to a chromatography column with a pipette, ensuring that no air bubbles get trapped in the adapter or gel bed.Resuspend cOmplete His-Tag Purification Resin by inverting the bottle several times.
Purifications with cOmplete His-Tag Purification Resin are compatible with various general purification formats including batch processing, column purification with gravity flow and automated processing using an automated chromatography system.Ĭolumn Packing Procedure for FPLC ApplicationsĬOmplete His-Tag Purification Resin is supplied as a 50% suspension in 20% ethanol. I would mention that the only issue is to be careful when you cure the white resin it can yellow off if you cure it too much.FPLC Purification Under Native Conditions Using the ÄKTAexplorer 100 SystemĪ detailed procedure for automated Fast Protein Liquid Chromatography (FPLC) using the ÄKTAexplorer 100 System is available in the Instructions for Use of cOmplete His-Tag Purification Column ( 06781535001, 06781543001), see chapter Purification Protocols > Purification Under Native Conditions.įPLC Purification Under Denaturing Conditions Using the ÄKTAexplorer 100 SystemĪ detailed procedure for automated Fast Protein Liquid Chromatography (FPLC) using the ÄKTAexplorer 100 System is available in the Instructions for Use of cOmplete His-Tag Purification Column ( 06781535001, 06781543001), see chapter Purification Protocols > Purification Under Denaturing Conditions. It works hard to make sure you are satisfied and are always happy to participate in community events. Now I know those numbers don't sound all that different, but when you print a model with 1,800 layers, shaving off a few seconds per layer makes a big difference. The Siraya Tech can give you the same quality print at just six to eight seconds and the first layer in 40-45 seconds. Most resin prints with an exposure time of around nine to 11 seconds, with a first layer exposure in 60-70 seconds. It's called "fast" resin for a good reason, too. It's perfect to create hollow prints since the resin maintains the part's strength without being brittle. Every print I have made with it so far has printed perfectly, and the resin seems to help smooth out layer lines that may appear. I have been using Siraya Tech Fast for ages now, and it's consistently the best resin I've used.