CLO3 | Caleosin-3

AS20 4418 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Product Information

Immunogen BSA-conjugated peptide, derived from N-terminus of Arabidopsis thaliana Caleosin 3, UniProt: O22788, TAIR: At2g33380

Host Rabbit

Clonality Polyclonal

Purity Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.

Format Liquid at 2 mg/ml.

Quantity 100 µg

Storage Store at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

Tested applications Western blot (WB)

Recommended dilution 1: 5000 (WB)

Expected | apparent MW 26,6 | 30 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity Brassica napus, Camelina sativa, Capsella rubella, Raphanus sativus Species of your interest not listed? Contact us

Not reactive in Poppulus sp.

Application examples

Application examples Western blot using anti-Caleosin antibodies

Arabidopsis thaliana senescent leaves were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE (crude extract) and denatured with 4X SDS buffer at 95°C for 5 min. 10 µg of protein was loaded/well and were separated on 15-20 % SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendations.

Additional information

Background

Background Caleosin3 encodes a calcium binding protein whose mRNA is induced upon treatment with NaCl, ABA and in response to desiccation. mRNA expression under drought conditions is apparent particularly in leaves and flowers. Isoform of caleosin with a role as a peroxygenase involved in oxylipin metabolism during biotic and abiotic stress. Involved in the production of 2-hydroxyoctadecatrienoic acid. The peroxygenase has a narrow substrate specificity thus acting as a fatty acid hydroperoxide reductase in vivo. Protective role to fungus pathogen has been indicaed. Expression is very low in young leaves and high in senescent leaves.

Product citations

Selected references Shimada et al. (2014). Leaf oil body functions as a subcellular factory for the production of a phytoalexin in Arabidopsis. Plant Physiol. 2014 Jan;164(1):105-18. doi: 10.1104/pp.113.230185.
Shimada et al. (2010). A rapid and non-destructive screenable marker, FAST, for identifying transformed seeds of Arabidopsis thaliana. Plant J. 2010 Feb 1;61(3):519-28. doi: 10.1111/j.1365-313X.2009.04060.x

calculated \| apparent molecular mass [kDa]:	26,6 \| 30 kDa
Clonality:	Polyclonal
Format:	Liquid at 2 mg/ml.
Host:	Rabbit
immunogen:	BSA-conjugated peptide, derived from N-terminus of Arabidopsis thaliana Caleosin 3, UniProt: O22788, TAIR: At2g33380
Purity:	Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
Quantity:	100 µg
recommended dilution:	1: 5000 (WB)
storage:	Store at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications:	Western blot (WB)
Picture (footer):	Arabidopsis thaliana senescent leaves were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE (crude extract) and denatured with 4X SDS buffer at 95°C for 5 min. 10 µg of protein was loaded/well and were separated on 15-20 % SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendations.
background:	Caleosin3 encodes a calcium binding protein whose mRNA is induced upon treatment with NaCl, ABA and in response to desiccation. mRNA expression under drought conditions is apparent particularly in leaves and flowers. Isoform of caleosin with a role as a peroxygenase involved in oxylipin metabolism during biotic and abiotic stress. Involved in the production of 2-hydroxyoctadecatrienoic acid. The peroxygenase has a narrow substrate specificity thus acting as a fatty acid hydroperoxide reductase in vivo. Protective role to fungus pathogen has been indicaed. Expression is very low in young leaves and high in senescent leaves.
Confirmed reactivity:	Arabidopsis thaliana
predicted reactivity:	Brassica napus, Camelina sativa, Capsella rubella, Raphanus sativus Species of your interest not listed? Contact us
not reactive in:	Poppulus sp.
All references:	Shimada et al. (2014). Leaf oil body functions as a subcellular factory for the production of a phytoalexin in Arabidopsis. Plant Physiol. 2014 Jan;164(1):105-18. doi: 10.1104/pp.113.230185. Shimada et al. (2010). A rapid and non-destructive screenable marker, FAST, for identifying transformed seeds of Arabidopsis thaliana. Plant J. 2010 Feb 1;61(3):519-28. doi: 10.1111/j.1365-313X.2009.04060.x