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Page 1: pI = (pk 1 + pk 2 )/2, the pH where the molecule has a net charge of 0
Page 2: pI = (pk 1 + pk 2 )/2, the pH where the molecule has a net charge of 0
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pI = (pk1 + pk2)/2, the pH where the molecule has a net charge of 0

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Phe-Ser-Tyr-Gly-Val-Gln

64 69

O2N

N

NO

HH

Phe

O

Val

OGln

HO

blue

green

green fluorescent protein (GFP)

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Ninhydrin Test

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Ion Exchange Chromatography

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Protein separation

size exclusion chromatography

ultrafiltration

ultracentrifugation

least soluble at their IP

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Step 1. Separation of Polypeptide Chains

dissociated by exposure to pH extremes, 8 M urea, 6 M guanidinium hydrochloride, or high salt cencentrations

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Cleavage of disulfide bridges

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Edman degradation

phenylisothiocyanate

50 cycles of reaction can be accomplished on 50 pmol (about 0.1 g)

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Carboxypeptidases cleave amino acid residues from the C-termini

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Cyanogen bromide

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Fmoc solid phase peptide synthesis: A Practical Approach

Edited by

Weng C. Chan and Peter D. White

OXFORD UNIVERSITYPRESS

2000

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