All-in-One cDNA Synthesis SuperMix-User Manual
Contents
1. biotool com Save time Save funding Poly FLAG Peptide lyophilized powder Description The Poly DYKDDDDK FLAG Peptide lyophilized powder was synthesized by 23 amino acid residues with molecular weight 2 864 Da The Asp Tyr Lys Xaa Xaa Asp motif is repeated three times in the peptide Eight amino acids at the C terminus make up the classic FLAG sequence Asp Tyr Lys Asp Asp Asp Asp Lys Components Properties Purityby HPLC MS by HPLC MS 95 33 aoa voren oncom is l 00 400 C trati mL for elute usion proteins from palea aN the Biotool Anti DYKDDDDK FLAG affinity gel Shipped in Bueice Bueice B Application For use in competitive elution of DYKDDDDK FLAG fusion proteins from the ANTI FLAG monoclonal antibody in solution or bound to agarose on the Biotool Anti DYKDDDDK FLAG affinity gel Notice 1 Read the User Manual carefully before the first use 2 Avoid freezing drying and high speed centrifugation during use and storage of beads 3 This product is for R amp D use only not for drug household or other uses Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices Procedure 1 Thoroughly suspend the Anti Flag Affinity Gel in the vial for a uniform suspension of the resin Quickly transfer 10ul of the gel suspension about 5ul of packed gel volume to a fresh tube 2 Add 0 6 mL TBS Thoroughly suspend the Anti Flag Affinity Gel by pi2. DK FLAG Affinity gel Wash the Biotool Antit DYKDDDDK FLAG Affinity gel three times with 5 mL of 50 glycerol with 10mM sodium phosphate 150 mM sodium chloride pH 7 4 containing 0 02 w v sodium azide Then add another 5 mL of 50 glycerol with 10 mM sodium phosphate 150 mM sodium chloride pH 7 4 containing 0 02 w v sodium azide and store at 20 C without draining Storage Store the product at 2 8 C Order amp Inquiry Tel 49 89 46148500 Fax 49 89 461485022 E mail eu order biotool com
3. petting Centrifuge the resin at 5000 rpm for 30 seconds and carefully remove the supernatant Be sure to remove all of the wash buffer without discarding the resin Repeat 3 4 times Order amp Inquiry Tel 713 732 2181 Fax 1 866 747 4781 E mail order biotool com 3 Add 500 uL of cell periplasmic extracts to the washed resin 4 Gently agitate samples for 2 hours at 4 C 5 Centrifuge the resin for 30 seconds at 5000 rpm Transfer the supernatants to a fresh tube 6 Wash the resin with 0 5mL TBS until the OD280 of the supernatant reads lt 0 05 7 Elution of DYKDDDDK FLAG Fusion Protein by Competition with Poly DYKDDDDK FLAG Peptide Elute the bound DYKDDDDK FLAG Fusion Protein by competitive elution with five one column volume aliquots of a solution containing 100 400 ug mL Poly DYKDDDDK FLAG Peptide in TBS 8 Recycle the Biotool Anti DYKDDDDK FLAG affinity gel Poly DYKDDDDK FLAG Peptide may not elute all of the DYKDDDDK FLAG Fusion Protein bound to Biotool Anti DYKDDDDK FLAG affinity gel It is recommended the Biotool Anti DYKDDDDK FLAG Affinity gel be regenerated immediately after use by washing with three 5 mL aliquots of 0 1 M glycine HCl pH 3 5 The gel should be immediately re equilibrated in TBS until the effluent is at neutral pH Note Do not leave the Biotool Antic DYKDDDDK FLAG Affinity gel in glycine HCI for longer than 20 minutes 9 Store the Biotool Anti DYKDDD
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