All-in-One cDNA Synthesis SuperMix-User Manual

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).
3. Add 500 µL 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<0.05.
Components
Contents
Cat. # B23111
Cat. # B23112
Size
4 mg
25 mg
Properties
Appearance (Form)
Form
Powder
Lyophilized Powder
Purity by HPLC-MS
Concentration
Shipped in
Storage Temperature
95.33 %
(Recommended working concentration is 100-400
µg/mL for elute FLAG fusion proteins from
the Biotool Anti-DYKDDDDK (FLAG) affinity gel.
Blue ice
2-8°C
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&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.
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 Anti-DYKDDDDK (FLAG) Affinity gel in
glycine HCl for longer than 20 minutes.
9. Store the Biotool Anti-DYKDDDDK (FLAG) Affinity gel
Wash the Biotool Anti-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.
Procedure
1. Thoroughly suspend the Anti-Flag Affinity Gel in the vial, for a
uniform suspension of the resin. Quickly transfer 10µl of the gel
suspension (about 5µl of packed gel volume) to a fresh tube.
2. Add 0.6 mL TBS. Thoroughly suspend the Anti-Flag Affinity Gel by
pipetting. 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 & Inquiry
Order & Inquiry
Tel: (713)732-2181
Fax: +1-866-747-4781
E-mail: order@biotool.com
Tel: +49-89-46148500
Fax: +49-89-461485022
E-mail: eu.order@biotool.com