Loading controls
In biology there are no universal controls. One has to determine a priori what does not change in one's test conditions and use that as a control. The choice of appropriate house-keeping protein would depend upon the particular experiment, and it needs to be confirmed. House-keeping proteins cannot be used for normalization if there is a suspicion that experimental conditions are affecting their expression.
Loading control guide
Other possible internal controls:
• Total protein (express fmol target protein/µg total protein)
• Dry mass
• Total chlorophyll (express fmol target protein/µg chlorophyll)
• Cells (express fmol target protein/cell)
• Total nitrogen (express fmol target protein/fmol N)
• Experiment specific ratios of particular proteins (ex. fmol PsbA/fmol RbcL)
Special notes:
FtsZ (AS07 217) can be used as a loading control in cyanobacteria.
GAPC1/2 use of this protein as a loading control should be supported with specific experimental data.
RbcL or RA (Rubisco activase) OEC223, AtpB (for Chlamydomonas reinhardtii, Rasala et al. 2013) or PsbO (OEE1) or PsbP (OEE2) for Chlamydomonas reinhardtii.
PsbO (OEE1) or PsbP (OEE2) are not so suitable for higher plants as accumulation of these proteins might drop to 12.5-25 % of the WT level in mutants defective for PSII core (Schult et al. 2007).
Tubulin alpha and gamma cannot be applied as a loading control for metal stress conditions. For more information check here.
AS16 4111 | Clonality: Monoclonal | Host: Mouse | Reactivity: Arabidopsis thaliana
AS13 2640 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Agostis stoloniferacv. ‘Penncross’,Arabidopsis thaliana, Brassica sp., Cannabis sativa L., Cucumis sativus, Cynara cardunculus, Glycine max, Hordeum vulgare, Nicotiana tabacum, Phoenix dactylifera, Setaria italica, Solanum tuberosum, Triticum aestivum, Zea mays
AS16 4111S | Protein/positive control
AS03 030 | Clonality: Polyclonal | Host: Chicken | Reactivity: [global antibody] for plant and bacterial F-type ATP synthases
AS05 085 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for plant, green alga, animal and bacterial F-type ATP synthases
AS16 3976 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Brassica oleracea var. botrytis cv. ’Diadom’, [compartment marker] of mitochondrial inner membrane
AS07 217 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Cyanobacteria
AS15 2894 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
AS10 710A | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, S. lycopersicum, V. faba, P. patens, S. europaea, Z. mays, Ch. reinhardtii | cellular [compartment marker] of nucleoplasm
AS15 2855 | Clonality: Polyclonal | Host: Chicken | Reactivity: A. thaliana, C. reinhardtii, L. esculentum, T. aestivum | cellular [compartment marker] of nucleoplasm
AS10 710 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. reinhardtii, H. vulgare, S. lycopersicum, V. faba, P. patens, S. europaea, Z. mays | cellular [compartment marker] of nucleoplasm
AS10 710-ALP | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, S. lycopersicum, V. faba, P. patens, S. europaea, Z. mays, Ch. reinhardtii | cellular [compartment marker] of nucleoplasm
AS10 710-HRP | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, S. lycopersicum, V. faba, P. patens, S. europaea, Z. mays, Ch. reinhardtii | cellular [compartment marker] of nucleoplasm
AS06 142-33 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A.thaliana, C. reinhardtii, C. vulgaris, C. sp. DT, E. crus-galli, F. arundinacea cv. Kord, H. pluvialis, H. spontaneum, H. vulgare, N. benthamiana, P. abies, P. banksiana, P. sativum, P. miliaceum, S. oleracea, S. tuberosum cultivar Taedong Valley, Synechococcus sp. PCC 7942 and PCC6803, T. salsuginea,
T.aestivum
compartment marker of thylakoid lumen
AS08 305 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, S. bicolor, Z. mays
AS06 142-23 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, Ch. reinhardtii, P. banksiana. , S.oleracea, T. aestivum
AS10 700 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. sativa, C. reinhardtii, H. spontaneum, F. pRatensis, G. max, G. hirsutum, G. barbadense, L. perenne, N. oceanica, N. tabacum, O. sativa, P. balsamifera, R. discolor, S. lycopersicum, Z. mays, T. salsuginea, red sulfur bacterium T. sp. Cad16 (isolated from Lake Cadagno)
AS01 017 | Clonality: Polyclonal | Host: Hen | Reactivity: [global antibody] for higher plants, algae, cyanobacteria | cellular [compartment marker] of plastid stroma
AS03 037 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for higher plants, lichens, algae, cyanobacteria, dinoflagellates, diatoms
compartment marker of chloroplast stroma in higher plants and cytoplasm in cyanobacteria
AS15 2832A | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
AS10 680 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. reinhardii, E. gracilis, H. vulgare, O. sativa, Setaria italica, Zea mays
AS10 681 | Clonality: Polyclonal | Host: Rabbit | Reactivity:Arabidopsis thaliana, Medicago sativa
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