Anti-EGFP sepharose Purification kit
Anti-EGFP sepharose, is consist of an EGFP VHH coupled to NHS-sepharose.
It is an efficient technique for isolating recombinant proteins or mammalian
expression proteins. The EGFP epitope system relies on a VHH EGFP recombinant antibody, which is able to react with N- and C- terminal GFP tagged and EGFP tagged fusion proteins. This product can be used for the immuneprecipitation or immune affinity purification.
The purified antibody is immobilized at 3-5 mg antibody per mL of 50% slurry
and this kit allows a rapid and efficient affinity purification of active EGFP fusion proteins. The affinity resin allows an efficient binding of EGFP fusion proteins without the need for preliminary steps and calibrations. The affinity bound EGFP fusion proteins can be efficiently eluted from the resin by acid condition. The eluted proteins can be used for characteristic analysis.
It is an efficient technique for isolating recombinant proteins or mammalian
expression proteins. The EGFP epitope system relies on a VHH EGFP recombinant antibody, which is able to react with N- and C- terminal GFP tagged and EGFP tagged fusion proteins. This product can be used for the immuneprecipitation or immune affinity purification.
The purified antibody is immobilized at 3-5 mg antibody per mL of 50% slurry
and this kit allows a rapid and efficient affinity purification of active EGFP fusion proteins. The affinity resin allows an efficient binding of EGFP fusion proteins without the need for preliminary steps and calibrations. The affinity bound EGFP fusion proteins can be efficiently eluted from the resin by acid condition. The eluted proteins can be used for characteristic analysis.
Component:
Note: Sepharose is 1:1 suspension in 20% ethanol as a preservative.
Product capacity:
The binding and elution capacity of 1 mL settled Anti-EGFP sepharose may be vary, commonly more than 1 mg GFP or EGFP fusion proteins. Trying different elution buffers for optimal results is recommended.
Materials Required but Not Provided:
For sustainable use and long term storage, store at 2 °C to 8 °C. DO NOT FREEZE.
Shipping Conditions:
Blue ice
| Reagent | Quantity for 10 rxns (C07009-K02) |
Quantity for 5 rxns (C07009-K01) |
composition |
| Anti-EGFP sepharose |
2 mL X 1 vial | 1 mL X 1 vial | 50% slurry of Anti-EGFP sepharose in 20% ethanol |
| Wash Buffer (10X concentration) |
5.0 mL X 1 vial | 2.5 mL X 1 vial | 10 mM Tris/Cl pH 7.5; 150 mM NaCl; 0.5 mM EDTA |
| Elution Buffer | 10 mL X 1 vial | 5 mL X 1 vial | 200 mM Glycine pH 2.5 |
| Neutralization Buffer | 2 mL X 1 vial | 1 mL X 1 vial | 2 M Tris pH 8.0 |
| spin column | 10 pcs | 5 pcs | |
| collection tube | 10 tubes | 5 tubes |
Product capacity:
The binding and elution capacity of 1 mL settled Anti-EGFP sepharose may be vary, commonly more than 1 mg GFP or EGFP fusion proteins. Trying different elution buffers for optimal results is recommended.
Materials Required but Not Provided:
- Micro-centrifuge capable of 15,000 x g
- 1.5 mL Centrifuge tubes
- End-over-end rotator
- CoIP Lysis Buffer (mild reaction): 10 mM Tris/Cl pH 7.5; 150 mM NaCl; 0.5 mM EDTA; 0.5% NP-40
- RIPA (vigorous reaction): 100 mM Tris/Cl pH 7.5; 300 mM NaCl; 0.2% Sodium Deoxycholate (or 0.1% SDS); 2% NP-40
- 2 x SDS Loading Buffer without DTT & 2 x SDS Loading Buffer with 1 mM DTT
- Pre-urea Buffer: 50 mM Tris pH 8.5; 1mM EGTA; 75 mM KCl
- Urea Elution Buffer: 6-8 M urea; 20 mM Tris pH 7.5; 100 mM NaCl
For sustainable use and long term storage, store at 2 °C to 8 °C. DO NOT FREEZE.
Shipping Conditions:
Blue ice