Suggested Control Antibodies for diatoms
AtpB | beta subunit of ATP synthase (10 µl) AS05 085-10 | clonality: polyclonal | host: rabbitThis antibody works well and detects both the chloroplastic and the mitochondrial forms of the enzyme. It usually gives a tight band and shows good reactivity. With ECL Advance detection we usually load 1 ug of total protein from diatom extracts, using this antibody.
RbcL | Rubisco large subunit, form I and form II (10 µl) AS03 037-10 | clonality: polyclonal | host: rabbit
This antibody is very 'hot' and the RUBISCO target is abundant, so only a small load of total protein is required to get a total band. With ECL Advance we usually load 0.5 or even 0.25 ug of total protein from diatom extracts, using this antibody. So depending upon the strength of the antibody to which you are comparing, this anti-RUBISCO may be too 'hot' to be a good comparison.
PsbA | D1 protein of PSII, C-terminal (10 µl) AS05 084-10 | clonality: polyclonal | host: rabbit
This antibody is used if the comparison protein is a hydrophobic membrane protein. This antibody works well but the PsbA protein is subject to fragmentation upon storage and freeze-thaw cycles. This might be a good index of the 'quality' of the total protein extracts. For this antibody we normally load 1 ug total protein, when detecting using ECL Advance.
Note: TMA-6 from Lumigen gives similar results to ECL Advance.
Protocol for protein extraction from diatoms.
Below you will find antibodies with confirmed and predicted reactivity to diatoms.
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AS08 300 | Extraction buffer for quantitative isolation of total soluble/membrane protein from plant tissue and algal/bacterial cells, optimized for subsequent western blot detection in denatured conditions
SA000002 | Clonality: Polyclonal | Host: Rabbit | Reactivity: higher plants and algae
AS05 070 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Dicots, monocots, algae
AS04 051 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Higher plants, algae, cyanobacteria
SA000003 | Clonality: Polyclonal | Host: Rabbit | Reactivity: higher plants, algae
AS09 409 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Dicots, monocots, algae, cyanobacteria
AS15 2992 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
AS03 030 | Clonality: Polyclonal | Host: Chicken | Reactivity: [global antibody] for plant and bacterial F-type ATP synthases
AS05 085-10 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for plant, green alga 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’
AS15 2991 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
AS10 935 | Clonality: Polyclonal | Host: Rabbit | Reactivity: diatoms
AS17 4116 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Heterosigma akashiwo
AS11 1789 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. acetabulum, A. thaliana, B. hypnoides (ulvophytes), G. theta, H. vulgare, Synechocystis sp., T. pseudonana, Z. mays
AS11 1789S | Protein standard for quantitation
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AS13 2671 Clonality: Polyclonal Host: Chicken Reactivity: [global antibody] for di- and monocots, conifers, ferns, mosses, green algae cellular [compartment marker] for plasma membrane
AS08 371S | Protein standard for quantitation and positive control
AS08 282 | Clonality:Polyclonal | Host:Rabbit | Reactivity:Diatoms
AS17 4115 | Clonality:Polyclonal | Host:Rabbit | Reactivity: Cyclotella sp., Thalassiosira pseudonana
AS01 021A | Clonality: Polyclonal | Host: Hen | Reactivity: [global antibody] for bacteria
AS08 310-100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, Ch. reinhardtii, red alga Gracilaria gracilis, diatom Thalassiosira sp. , P. tricornutum Bohlin, P. yunnanensis Dode, S. tuberosum
AS08 310 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, C. reinhardtii, red alga Gracilaria gracilis, diatom Thalassiosira sp., P. tricornutum Bohlin, P. yunnanensis Dode, S. lycopersicum, S. tuberosum
AS09 458S | Protein standard for quantitation and positive control
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AS09 458 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. comosus, A. thaliana, C. ciliaris, C. gayana, C. velia, H. vulgare, J. curcas, L. fusca, Lupinus sp. , M. maximus, M. crystallinum, N. tabacum, O. sativa, P. antidotale, P. coloratum, P. strobus, Saccharum spp. hybrid clone C91-301, S. lanata, S. laricifolia, S. bicolor, Synechocystis PCC 6803, Phaeodactylum tricornutum (strain CCAP 1055/1), T. weissfloggi, Z. mays, Z. muelleri
AS09 458-ALP | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. comosus, A. thaliana, S. PCC 6803, Ch. velia, H. vulgare, J. curcas, Lupinus sp. , M. crystallinum, N. tabacum, O. sativa, Saccharum spp. hybrid clone C91-301, S. laricifolia, Z. mays, Synechocystis sp. , P. tricornutum (strain CCAP 1055/1), T. weissfloggi
AS09 458-HRP | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. comosus, A. thaliana, S. PCC 6803, Ch. velia, H. vulgare, J. curcas, Lupinus sp. , M. crystallinum, N. tabacum, O. sativa, Saccharum spp. hybrid clone C91-301, S. laricifolia, Z. mays, Synechocystis sp. , P. tricornutum (strain CCAP 1055/1), T. weissfloggi
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