TapC2

TapC2 was made by PCR from human Tap1, residues 550 (methionine) to 619 (C-terminus). Cloned into pMW172 NdeI/EcoRI.

Growth

  1. Transform BL21 DE3 [RIL] with pMW172/TapC2 DNA. Select on Amp plates.
  2. Pick a single colony into 1 l 2xTY/Amp. Grow overnight at 37°C with shaking.
  3. Harvest by centrifugation at 5,000 rpm at 4°C for 10 minutes.
  4. Resuspend in 10 ml sucrose buffer per litre of media. Store at -20°C.

Purification

  1. EXCLUSION BODIES

refolding

Quantification

Coefficient at 280 nm = 12,660 M-1 cm-1.

Mr = 12,566.

Enter
A280 reading 		Enter
dilution factor 		  Conc (mM) Conc (mg ml-1)

Buffers

HiLo HiHi
20 mM Tris-Cl pH 8.5
1 mM EGTA
1 mM DTT
 		 20 mM Tris-Cl pH 8.5
1 mM EGTA
1 mM DTT
500 mM NaCl
RPG 2000/08/25

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