Loading controls

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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 suspcion that experimental conditions are affecting their expression.

Other possible internal controls:
- total protein (express fmol target protein/ug total protein)
- dry mass
- total chlorophyll (express fmol target protein/ug 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)

For short term experiments Rubico protein makes a good loading control as its turnover is slow. However, Rubisco level do vary with nutrient levels, particularly nitrogen.

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).

Below you will find proposed antibodies to use as a loading control.

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ACT | Actin

272 €

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AS13 2640 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. stolonifera cv. ‘Penncross’, A. thaliana, C. sativus, G. max, H. vulgare, N. tabacum, S. lycopersicum, S. tuberosum, Z. mays

Actin-11

272 €

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AS10 702 | Clonality: Monoclonal | Host: Mouse | Reactivity: A. thaliana, N. cataria, N. rtanjensis, N. tabacum, O. sativa, S. tuberosum, Z. mays

AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (chicken)

355 €

AS03 030 | Clonality: Polyclonal | Host: Chicken | Reactivity: [global antibody] for plant and bacterial F-type ATP synthases

AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (rabbit)

272 €

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AS05 085 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for plant, green alga, animal and bacterial F-type ATP synthases

AtpB | Beta subunit of ATP synthase, mitochondrial

272 €

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AS16 3976 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Brassica oleracea var. botrytis cv. ’Diadom’

FtsZ | Procaryotic cell division GTPase (cyanobacterial)

360 €

AS07 217 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Cyanobacteria

GAPC1/2 | Glyceraldehyde-3-phosphate dehydrogenase

272 €

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AS15 2894 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

H3 | Histone H3 (100 µl)

407 €

AS10 710-100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, S. lycopersicum, V. faba, P. patens, S. europaea, Z. mays, Ch. reinhardtii | cellular [compartment marker] of nucleoplasm

H3 | Histone H3 (affinity purified)

272 €

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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

H3 | Histone H3 (chicken antibody)

272 €

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AS15 2855 | Clonality: Polyclonal | Host: Chicken | Reactivity: A. thaliana, C. reinhardtii, L. esculentum | cellular [compartment marker] of nucleoplasm

H3 | Histone H3 (rabbit antibody) (nuclear marker)

272 €

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AS10 710 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. reinhardtii, S. lycopersicum, V. faba, P. patens, S. europaea, Z. mays | cellular [compartment marker] of nucleoplasm

H3 | Histone H3, ALP-conjugated (40 µg)

417 €

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

H3 | Histone H3, HRP-conjugated (40 µg)

417 €

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

PsbO | 33 kDa of the oxygen evolving complex (OEC) of PSII (anti-protein)

272 €

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AS06 142-33 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A.thaliana, C. reinhardtii, C. sp. DT, F. arundinacea cv. Kord, H. pluvialis, H. spontaneum, H. vulgare, N. benthamiana, P. abies, P. banksiana, S. oleracea, S. tuberosum cultivar Taedong Valley, Synechococcus sp. PCC 7942 and PCC6803, T. salsuginea, T.aestivum

PsbP | 23 kDa protein of the oxygen evolving complex (OEC) of PSII (anti-monocot protein)

355 €

AS08 305 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, S. bicolor, Z. mays

PsbP | 23 kDa protein of the oxygen evolving complex (OEC) of PSII (anti-protein)

355 €

AS06 142-23 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, Ch. reinhardtii, P. banksiana. , S.oleracea, T. aestivum

RA | Rubisco activase

272 €

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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)

RbcL | Rubisco large subunit, form I and form II

265 €

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AS03 037 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for higher plants, licHens, algae, cyanobacteria, dinoflagellates, diatoms | cellular [compartment marker] of plastid stroma in higher plants and cytoplasm in cyanobacteria

RbcL | Rubisco large subunit, form I

355 €

AS01 017 | Clonality: Polyclonal | Host: Hen | Reactivity: [global antibody] for higher plants, algae, cyanobacteria | cellular [compartment marker] of plastid stroma

RPN6 | 26S proteasome non-ATPase regulatory subunit 9

272 €

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AS15 2832A | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

RPN6 | 26S proteasome non-ATPase regulatory subunit 9 (A.thaliana)

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AS15 2845 | Clonality: polyclonal | Host: rabbit | Reactivity: Arabidopsis thaliana

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Tubulin alpha chain

272 €

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AS10 680 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. reinhardii, E. gracilis, H. vulgare, O. staiva, Zea mays

Tubulin beta chain

381 €

AS10 681 | Clonality: Polyclonal | Host: Rabbit | Reactivity:Arabidopsis thaliana, Medicago sativa