Can I use the IEF Marker, 3-10 for protein quantitation?
The exact amount of each protein in the IEF Marker, 3-10 is not known. We do not recommend using this marker for quantitation purpose.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
Can I use the IEF Marker 3-10 with IPG strips?
The IEF Marker, 3-10 is not designed for use with IPG strips.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
What are the isoelectric points (pIs) and origins of the proteins in the IEF Marker, 3-10?
The IEF Marker 3-10 is a ready-to-use, unstained protein marker especially developed for IEF applications. It contains 13 purified proteins ranging in pI from 3-10. It works well on horizontal and vertical one-dimensional IEF gels and can be used for both native and denaturing conditions (e.g., 8 M urea, to determine the isoelectric point (pI) of unknown protein samples. Here are the pIs and origins of the proteins in the IEF Marker, 3-10:
- Band no. 9: Cytochrome C; Pig heart origin; 10.7 pI
- Band no. 8 Ribonuclease A; Bovine pancreas origin; 9.5 pI
- Band no. 7 c, m, a Lectin; Lens culinaris origin; 8.3, 8.0, 7.8 pI
- Band no. 6 c, a Myoglobin; Horse muscle origin; 7.4, 6.9 pI
- Band no. 5 Carbonic anhydrase; Bovine erythrocytes origin; 6.0 pI
- Band no. 4 c, a B-Lactoglobulin; Bovine milk origin; 5.3, 5.2 pI
- Band no. 3 Trypsin inhibitor; Soybean origin; 4.5 pI
- Band no. 2 Glucose oxidase; Aspergillus niger origin; 4.2 pI
- Band no. 1 Amyloglucosidase; Aspergillus niger origin; 3.5 pI
Note: Temperature = 5 degrees C for pI values given. pI is temperature dependent, i.e. decreases as temperature increases (approximately -0.005 to-0.03 pH units per degree C)
c = cathodic; m = middle; a = anodic
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
What are the storage conditions and shelf life for the IEF Marker 3-10?
We recommend storing the IEF Marker 3-10 at -20 degrees C and it is stable for 1 year when properly stored.
Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.
My monoclonal antibody runs as a smear on an IEF gel, while the IEF marker focuses perfectly. Why is this?
It is not unusual for antibodies (even monoclonals) to be differentially glycosylated, and therefore not focus well on IEF gels. Sometimes a monoclonal can even run more smeary or unfocused than a polyclonal, over a range of pH 6.5 to 8.0. Try running the gel longer to improve the focus, but often the improvement may be minimal if any at all.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
