Anti-mCherry sepharose Purification kit
Anti-mCherry sepharose, is consist of an mCherry VHH coupled to NHS-sepharose. It is an efficient technique for isolating recombinant proteins or mammalian expression proteins. The mCherry epitope system relies on a VHH mCherry recombinant antibody, which is able to react with N- and C- terminal RFP, DsRed and mCherry tagged fusion proteins. This product can be used for the immuneprecipitation or immune affinity purification.
The purified antibody is immobilized at 2-4 mg antibody per ml of 50% slurry
and this kit allows a rapid and efficient affinity purification of active mCherry
fusion proteins. The affinity resin allows an efficient binding of mCherry fusion
proteins without the need for preliminary steps and calibrations. The affinity
bound mCherry fusion proteins can be efficiently eluted from the resin by acid
condition. The eluted proteins can be used for characteristic analysis.
The purified antibody is immobilized at 2-4 mg antibody per ml of 50% slurry
and this kit allows a rapid and efficient affinity purification of active mCherry
fusion proteins. The affinity resin allows an efficient binding of mCherry fusion
proteins without the need for preliminary steps and calibrations. The affinity
bound mCherry 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-mCherry sepharose may be vary, commonly more than 1 mg RFP, DsRed or mCherry 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 (C07011-K02) |
Quantity for 5 rxns (C07011-K01) |
composition |
| Anti-mCherry sepharose |
2 mL X 1 vial | 1 mL X 1 vial | 50% slurry of Anti-mCherry sepharose in 20% ethanol |
| Wash Buffer (10X concentration) |
5.0 mL X 1 vial | 2.5 mL X 1 vial | 100 mM Tris/Cl pH 7.5; 1.5 M NaCl; 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-mCherry sepharose may be vary, commonly more than 1 mg RFP, DsRed or mCherry 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