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FUM1 + FUM2 | Fumarase 1+2 (chloroplastic+mitochondrial)

AS16 3966 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

FUM1 + FUM2 | Fumarase 1+2 (chloroplastic+mitochondrial) in the group Antibodies for Plant/Algal  / Carbohydrates at Agrisera AB (Antibodies for research) (AS16 3966)

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Product Information

Immunogen KLH-conjugated peptide chosen from Arabidopsis thaliana FUM protein sequence, mitochondrial (FUM1): P93033, TAIR: At2g47510, chloroplastic (FUM2): UniProt Q9FI53
TAIR: At5g50950
Host Rabbit
Clonality Polyclonal
Purity Serum
Quantity 50 µl
Reconstitution

 for reconstitution add 50 µl of sterile water

Storage The antibody may be stored at -20℃for one year in its original formulation. Additionally, antibody may be stored at 2℃ to 8℃ for up to 1 month without detectable loss of activity. Avoid repeated freeze-thaw cycles of the diluted antibody.
Tested applications Western blot (WB)
Recommended dilution

1:1000 (WB)

Expected | apparent MW

49.9 | 55 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Ananas comosus, Capsicum chinense, Cynara cardunculus var. scolymus, Genlisea aurea, Glycine soja, Gossypium hirsutum, Helianthus annuus, Hordeum vulgare, Nicotiana sylvestris, Nicotiana tabacum, Oryza sativa, Rhizophora mucronata, Ricinus communis, Solanum lycopersicum, Trifolium pratense, Zea mays, Vigna radiata, Quercus suber

Application examples

Application examples Application example


Western blot using anti-FUM antibodies


Total protein extracted freshly from Arabidopsis thaliana 14-old-day seedlings grown on ½ MS without sugar in extraction buffer [100 mM NaCl; 50 mM Tris–HCl pH 7.5; 0.5% (v/v) Triton X-100; 1 mM DTT; 1× Complete Protease Inhibitor Cocktail Tablet (Roche, Bassel, Switzerland)] and denatured in Laemeli buffer at 55°C for 5 min. 50 µg of protein extraccts: Arabidopsis thaliana wt (1), fum2-1 mutant (2), fum2-1 + GFP-FUM2 N1 (3), fum2-1 + GFP-FUM2 N2 (4), fum2-1 + GFP-FUM2 N3 (5), fum2-1 + GFP-FUM2 N3 (6), no protein -GFPfum2-2 (8) were loaded and separated on 12% SDS-PAGE and blotted 1h to PVDF membranes in a semi-dry transfer system. The membrane was blocked with 3% milk for 1h at RT with agitation and then incubated in the primary antibody (anti-FUM) at a dilution of 1: 2 000 for 1h at RT with agitation in TBS-T and milk 3%. The antibody solution was decanted and the blot was rinsed briefly three times in TBS-T for 5 min. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:5 000 in for 1h at RT with agitation. The blot was washed as above and developed for 10 min using chemiluminescnce detection reagent in a Gel Doc XR+ System (Bio-rad; Hercules; USA).

fum1 mutant is lethal therefore a band of ca. 60 kda is still present in fum2-1 and fum2-2 background.

Courtesy of Dr. Antoni Garcia Molina, Ludwig-Maximilians-Universität München (LMU), Germany

Additional information

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antibodies to proteins involved in carbohydrate metabolism

Background

Background FUM (Fumarase) Fumarate hydratase 1, mitochondrial (FUM1), Fumarate hydratase 2, chloroplastic (FUM2) are enzymes which belong to a subpathway of tricarboxylic acid cycle, part of carbohydrate metabolism.

Product citations

Selected references

To be added when available, antibody released in October 2018.

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