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ELISA
SDS-PAGE
Bioactivity
the solution can be further dialyzed against PBS (phosphate buffered saline
10 mM Tris-HCl
2 mM dithiothreitol
Enzyme Activity
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. Dulbecco's PBS).
ELISA.
Western Blot.
Flow Cyt
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.
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
e.g. Dulbeccos PBS).
e.g. Dulbeccos PBS).
See References 2 and 3 for more details.
pH 7.4).
For immunization purposes
pH 7.4).
For Immunization purposes
Biological activity/ Working concentration:
Optimum concentration for human umbilical vein endothelial cells (HUVEC) range from 50-200 μg/ml
ELISA.
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)
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
Can be used as standard in a
Sandwich ELISA.
ELISA.
Western blotting.
FACS
Protein standard in 1D and 2D SDS gel electrophoresis.
Immunoassay.
Antigen for immunization.
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
Secreted
Suitable for Immunoblotting (Western or Dot blot)
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.Dulbecco's PBS).
et al. (1991) Biochem. Pharmacol. 42: 1661.
3. Satoh T.
et al.(1992) J.Biol.Chem. 267: 2537.
4. Weiss R.
optimal concentration with heparin (30 μg/ml) is about 12 μg/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
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
000-1/32
000-1/40
000.
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: 1/500-1/2
000.
Western blot: 1/10
500.
Immunohistochemistry: 1/200-1/1
ELISA.
Western Blot.
Cell culture and/or animal studies.
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.
Western blotting.
ELISA:
this protein is suitable for use as a Mouse IgE standard in ELISA assays.
ELISA
.
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 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.
Functional Assays.
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.
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
Specific methodologies have not been tested using this product.
Suitable for Immunoblotting (Western or dot blot)
Western Blot.
ELISA.
Western Blot.
Protocol: 1. Lauber T
Western Blotting.
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 (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
Pathways
×
Olfactory transduction
Protein Source
×
__Category ID
×
Assay Platforms
×
C-Tag
×
Mammalian Cell Selection
×
Neomycin
Puromycin
None
Recombinant
×
__Matched ELISA Pair
×
Carrier-free
×
__PT
×
1
2
Biosimilar
×
Fluorescent Label
×
Viral Packaging
×
Secretory
×
Citations
×
Yes
Availabilities
×
In Stock
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