Native Proteins

Proteins

Native Proteins

Druggable Genome

10 mM Tris-HCl

2 mM dithiothreitol

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoassays.
Immunization.

Secreted Protein

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

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

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).
See References 2 and 3 for more details.

pH 7.4).
For Immunization purposes

Protease

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

Protein standard in 1D and 2D SDS gelelectrophoresis.
Immunoassays.
Immunization.
Protocol: Reconstitution to Filaments is performed by dissolving in 6 M urea buffer (see above) at concentrations of approx. 0.5 mg/ml. 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

Protein standard in 1D and 2D SDS gel electrophoresis.
Immunoassay.
Antigen for immunization.

Suitable for Immunoblotting (Western or Dot blot)

Transmembrane

e.g. Dulbeccos PBS).

pH 7.4).
For immunization purposes

000-1/32

000.

000.
Western Blot: 1/500-1/2

500.
Immunohistochemistry: 1/200-1/1

ELISA.
Suitable for use as a Control or standard in indirect trapping ELISA for quantitation of antigen in serum using a standard curve.
Immunoprecipitation.
Western blotting.

ELISA.
Western Blot.

ELISA: this protein is suitable for use as a Mouse IgE standard in ELISA assays.

ELISA.

ELISA and Immunohistochemistry as well as other beta galactosidase-avidin/biotin based enzymatic assays. This product has been assayed against 1.0 µg of biotinylated IgG in a standard capture ELISA using OPNG as a substrate for 30 minutes at room temperature.
Recommended Dilutions:
ELISA: 1/8000-1/32000.
Western Blot: 1/500-1/2500.
Immunohistochemistry: 1/200-1/1000.

ELISA and immunohistochemistry as well as other glucose oxidase-antibody based enzymatic assays.
Recommended Dilutions:
ELISA: 1/8

ELISA.

ELISA.
Western Blot.

ES Cell Differentiation/IPS

For use as a Control or Standard in Indirect trapping ELISA for quantitation of antigen in serum using a standard curve

For use as a Control or standard in indirect trapping ELISA for quantitation of antigen in serum using a standard curve.
Immunoprecipitation.
Western blotting.

Specific methodologies have not been tested using this product.

change growth medium against basal medium (EBM) in the early afternoon]
- change EGF against EBM (for HUVEC: EBM +1-2% FCS)
- incubate 24 h
- change medium again and add factors (growth factors

etc.)
- incubate for 18 h
- add 10 µl 3H-Thymidine solution [0.025 mCi/ml] per well (=0.25 µCi)
- incubate another 6h at 37°C
- Washing steps: (250 µl/well)
PBS 1x
MeOH 2x 5 min
TCA 2x 10 min
H2O 1x
- lyse cells in 250 µl 0.3M NaOH per well
- transfer 2.5 ml ECO Plus into the appropriate scintillation vials
- transfer cell lysats into the scintillation vials
- count by liquid scintillation (ß-counter; Beckmann Instruments)

for immunoprecipitation and for Wwestern blotting.

inhibitors

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

optimal concentration with heparin (50 μg/ml) is about 12 μg/ml.
Protocol: Thymidine Incorporation with HUVEC
- plate cells with a density at 5-7 x 10e3 cells/well in growth medium (EGM)
- incubate cells over night [if urgent

plate cells in the morning

Brain

2