nativemarker

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Native - Standards

April 28, 2023

These standards cannot be used to determine the mol-weights, the electrophoretic migration-speed of the proteins are also related to their charge within the used buffer-system; Q/MW, where Q is the charge and MW the mol-weight.

Pattern in native electrophoresis are compared to other (known) patterns to determine the proteins.
These markers are used to differ between problems in electrophoresis and problems with the samples themselves!

Dilutions* of the markers are given when as a visualization process Coomassie-staining is chosen.
Fluorescence: dilute 10 + 30. For silver-staining dilute 10 + 90

Product	dilution* cathodal native (marker + gel buffer)	dilution* anodal native (marker + gel buffer)	Prestained ?	Loading (per band)	Order Number
VWR: HMW 66-669 kD	---	400 ul buffer per bottle	---	~380 ng in 15 ul	95016-850
VWR: IEF 3-10	250 ul buffer per bottle	---	---	~600 ng in 15 ul	95016-832
Sigma: Ultra Low Range 1,060-26,600	30 ul + 570 ul	30 ul + 770 ul	---	?	M3546
Native Protein-Mix (see below)	50 ul + 150 ul	50 ul + 150 ul	Myoglobin, Cytochrom C	500 ng in 15 ul	- self mix -

Native Protein-Mix: each 0,25 mg/ml. Can be used in both directions, anodal and cathodal. Should look like GE’s IEF Mix 3-10.
Amyloglucosidase - pI 3.5, Glucose oxidase - pI 4.2, Trypsin inhibitor - pI 4.5, β-Lactoglobulin - pI 5.15 / 5.3
Carboanhydrase - pI 6.0, Myoglobin horse - pI 6.9 / 7.35, Lentil lectin - pI 7.75 / 8.0 / 8.3, Ribonuclease A - pI 9.45
Cytochrome C - pI 10.65. Sample volume 5 ul.
See special application: Kathodische native Elektrophorese.... (S.Dammeier, Tübingen)


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	Native gel 10%T, special buffer pH 7.4 (anodal run), Hot Coomassie stained


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	4

		3

	2

1


	Native gel 15%T, spacial acicic buffer, (cathodal run), Hot Coomassie stained


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		-


	3

		2

		1


	Standard protein mix (cathodal run) (S.Dammeier)


	EDC Electrophoresis Development & Consulting, Vor dem Kreuzberg 17, 72070 Tübingen (Germany)