Recombinant Proteins

Proteins

Recombinant Proteins

Druggable Genome

Transmembrane

Secreted Protein

Transcription Factors

Protein Kinase

ES Cell Differentiation/IPS

Protease

Stem cell - Pluripotency

Phosphatase

Adult stem cells

Ion Channels: Other

Embryonic stem cells

Nuclear Hormone Receptor

P450

Cancer stem cells

GPCR

Induced pluripotent stem cells

Stem cell relevant signaling - JAK/STAT signaling pathway

Stem cell relevant signaling - DSL/Notch pathway

Stem cell relevant signaling - TGFb/BMP signaling pathway

ELISA

Stem cell relevant signaling - Wnt Signaling pathway

SDS-PAGE

Ion Channels: Potassium

Bioactivity

Ion Channels: Glutamate Receptors

the solution can be further dialyzed against PBS (phosphate buffered saline

Ion Channels: ATP Receptors

Ion Channels: Cys-loop Receptors

10 mM Tris-HCl

2 mM dithiothreitol

Enzyme Activity

e.g. Dulbecco's PBS).

Flow Cyt

ELISA.
Western Blot.

Ion Channels: Transient receptor potential

Ion Channels: Sodium

both dissolved in 9.5 M urea buffer (see above). Protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4 M urea and then to low salt condition (50 mM NaCl

pH 7.4). For immunization purposes

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoassays.
Immunization.
Protocol: Reconstitution to filaments is performed by mixing equimolar amounts of keratins of type I and type II at concentrations of approx. 0.5 mg/ml

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoassays.
Immunization.
Protocol: Reconstitution to filaments is performed by mixing equimolar amounts of keratins of type I and type II at concentrations of approx. 0.5 mg/ml

pH 7.4).
For immunization purposes

pH 7.4).
For immunization purposes

Ion Channels: Cyclic nucleotide gated

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoblotting.
Receptor-binding studies.

Western Blot.

0.1% (w/v) insulin

000-1/5

10 mM sodium phosphate

10mM Tris-HCl

1mM DTT

2 mM dithiothreitol

2mM EDTA.
2. Add 4ug of recombinant thioredoxin into reaction cocktail and mix immediately.
3. Incubate at 25°C for 20 minutes.
4. Record the increase in A650nm for 15 minutes

2mM dithiothreitol

63mM sodium phosphate

Biological activity/ Working concentration: The working concentration of ECGF for HUVEC is in the range of 25µg/ml to 100µg/ml. When adding Heparin (2.5mg per mg ECGF) an ECGF concentration of 12µg/ml (30µg/ml Heparin) is optimal. In this case 6mg ECGF are sufficient for 500 ml medium.

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoassays.
Immunization.
Protocol: Reconstitution to filaments: After vimentin is dissolved in 9.5M Urea buffer (see above)

Can be used as standard in a Sandwich ELISA.

ELISA.

FACS

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoassays.
Immunization.
Protocol: Reconstitution to filaments: after desmin is dissolved in 9.5 M urea buffer (see above)

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoassays.
Immunization.
Protocol: Reconstitution to filaments: after vimentin is dissolved in 6 M urea buffer (see above)

Protocol: 1. Uehara Y.

Protocol: Activity Assay
1. Prepare a 1 ml reaction cocktail into a suitable container: The final concentrations are 5mM Tris

and Fukazawa H. (1991) Methods Enzymol. 201: 370.
2. Fukazawa H.

and Nuccitelli R. (1992) J.Biol.Chem. 267:5608.

both dissolved in 9.5 M urea buffer (see above).
Protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4 M urea and then to low salt condition (50 mM NaCl

e.g. Dulbeccos PBS).

e.g.Dulbecco's PBS).

et al. (1991) Biochem. Pharmacol. 42: 1661.
3. Satoh T.

et al.(1992) J.Biol.Chem. 267: 2537.
4. Weiss R.

protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4 M urea and then to low salt condition (50 mM NaCl

protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4M Urea and then to low salt condition (50mM NaCl

protofilaments and filament complexes are obtained by dialyzing the resulting polypeptide solution stepwise to a concentration of 4M urea and then to low salt condition (50 mM NaCl

the solution can be further dialyzed against PBS (Phosphate Buffered Saline

000) as well as other antibody based fluorescent assays.

000)
Flow Cytometry/FACS (1/1

000-1/200

000-1/40

000.
Immunohistochemistry: 1/1

000.
Protein G Peroxidase is a useful reagent in Western Blotting and ELISA experiments. Protein G Peroxidase can be utilized as a pseudo-secondary detection reagent when used in conjunction with an IgG-based primary antibody and appropriate substrate (such as TMB-1000 or Femtomax-110).

000.
Western blot: 1/10

Biological activity/ Working concentration: Optimum concentration for human umbilical vein endothelial cells (HUVEC) range from 50-200 μg/ml

ELISA.
Western Blot.
Cell culture and/or animal studies.

ELISA.
Western blotting.

Functional Assays.

Positive control for Western blot analysis

Positive control for Western blot analysis.
Standard for ELISA.

Adermann K. Role of the prosequence of guanylin. Protein Sci. 1999 Sep; 8(9): 1850-9.
4. Lauber T

Cell culture and/or animal studies.
ELISA.
Western Blot.

Cohen MB. Proguanylin secretion and the role of negative-feedback inhibition in a villous epithelial cell line. Am J Physiol Gastrointest Liver Physiol. 2002 Sep; 283(3): G695-702.
3. Schulz A

ELISA.
Functional Assays.

ELISA.
Western blotting.

Forssmann WG

Hawkins JA

Hidaka Y

Immunofluorescence Microscopy (1/1

Immunoperoxidase Electron Microscopy and Immunohistochemistry as well as other peroxidase-biotin-avidin based enzymatic assays.
Recommended Dilutions:
ELISA: 1/20

Marx UC

Marx UC. Native and recombinant proguanylin feature identical biophysical properties and are monomeric in solution. Biochemistry. 2002 Dec 10; 41(49): 14602-12.
2. Rudolph JA

Marx UC. Solution structure of human proguanylin: the role of a hormone prosequence. J Biol Chem. 2003 Jun 27; 278(26): 24118-24. Epub 2003 Apr 21.

Neudecker P

Nourse A

Rosch P

Schulz A

Shimonishi Y

Suitable for Immunoblotting (Western or dot blot)

Western Blot.
ELISA.

Western Blot.
Protocol: 1. Lauber T

Western Blotting.

optimal concentration with heparin (30 μg/ml) is about 12 μg/ml.

2