Waters RapiGest SF Surfactant - Manual

[ Care and Use ManUal ]
RapiGest SF Surfactant
I. IntroductIon
RapiGest ™ SF is a reagent used to enhance in-gel (Ref. ) and in-solu-
tion (Refs. 2-6) enzymatic digestions of proteins. Recent investigations
also support its use to solubilize glycoproteins prior to deglycosylation
with enzymes such as PNGase F (Publication in preparation). RapiGest
SF helps solubilize proteins making them more susceptible to enzy-
matic cleavage without significantly inhibiting enzyme activity. Unlike
other commonly used denaturants (e.g., SDS or Urea), RapiGest SF
does not modify peptides or suppress endoprotease activity. RapiGest
SF is compatible with enzymes such as Trypsin, Lys-C, Asp-N and Glu-
C and other enzymes. This reagent is easily removed after use allowing
MALDI-TOF MS, HPLC/MS, or HPLC/UV analyses of digested samples.
II. Storage and StabIlIty
The lyophilized powder is stable at room temperature until the expiration
date printed on the label (i.e., 3 years after packaging). Once reconsti-
tuted in high purity water or a buffer (pH 7–0) the solution is stable
for one week when stored at 2–8 °C. Long term storage of frozen ali-
quots is possible but not recommended due to potential solubilization
issues of RapiGest SF or storage buffer.
Note: RapiGest SF hydrolyzes in acidic solutions (half life 8 min. at pH
2 and 60 min. at pH 3).
III. reconStItutIon of rapIgeSt Sf powder
Shown in Table , are the volumes required to reconstitute the mg
RapiGest SF powder in water or buffer to obtain preferred concentra-
tions. If Mass Spec analysis is required, the recommended buffer is
50mM Ammonium Bicarbonate (NH
4
HCO
3
). Alternative buffers, such
as 0 mM Tris-HCl or 25 mM sodium phosphate, are also RapiGest SF
compatible.
contentS
I. IntroductIon
II. Storage and StabIlIty
III. reconStItutIon of RapigeSt Sf powder
IV. SuggeSted procedure for In-SolutIon dIgeStIonS
V. SuggeSted procedure for In-gel dIgeStIonS
VI. SuggeStIonS for worKIng wItH proteolytIc reSIStant
or HydropHobIc proteInS
VII. SuggeSted SaMple preparatIon for Hplc and MaSS
SpectroMetry
RapigeSt Sf Surfactant