SAG12 | Senescence-specific cysteine protease SAG12

AS14 2771 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, B. napus

Product Information

Immunogen

KLH-conjugated peptide derived from Arabidopsis thaliana SAG12 protein sequence, UniProt:Q38886, TAIR: AT5G45890

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized in PBS pH 7.4.

Quantity 50 µg

Reconstitution For reconstitution add 50 µl of sterile water.

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

Tested applications Western blot (WB)

Recommended dilution 1 : 2000 (WB)

Expected | apparent MW

38 | 28 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Brassica napus, Hordeum vulgare

Predicted reactivity Actinidia deliciosa, Brassica rapa, Gossypium hirsutum, Petunia hybrida, Morus notabilis, Nicotiana tabacum, Petunia hybrida, Populus trichocarpa, Ricinus communis, Theobroma cacao
Species of your interest not listed? Contact us

Not reactive in Citrus sinensis

Application examples

Application example

20 - 1.25 µg of soluble proteins from senescing or young leaves of oilseed rape (Brassica napus L.) extracted with citrate sodium-phosphate buffer (100 mM, pH 6.8) and were separated on 12% SDS-PAGE and blotted 10 min to PVDF using semi-dry transfer (see Desclos et al. 2009). Blots were washed with TBS-T 3 times for 15 min at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 (prepared in TBS-T with 5% low fat milk) for 2h or 1h30min at RT with agitation. The antibody solution was decanted and the blot was washed 5 times for 5 min in TBS-T and 3 times for 5 min in TBS at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:0 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 1 second.

Desclos M, Etienne P, Coquet L, Jouenne T, Bonnefoy J, Segura R, Reze S, Ourry A, Avice J-C. 2009. A combined 15N tracing/proteomics study in Brassica napus reveals the chronology of proteomics events associated with N remobilisation during leaf senescence induced by nitrate limitation or starvation. Proteomics 9, 3580-3608.

Courtesy of Dr. Jean-christophe Avice, INRA, France

Additional information

Recommended plant extraction buffer: See Durian et al. (2019). Denature in Laemmli-sample buffer (final concentrations: 5% (v/v) beta-mercaptoethanol, 69 mM Tris-HCl pH 6.8, 11.1% (v/v) glycerol, 2.15 % (v/v) SDS) at 70◦C for 5 min

Related products

AS16 3110 | Anti-Cp2 | Cysteine protease, rabbit antibodies

collection of antibodies involved in senescence

Algal protein extraction buffer

Secondary antibodies

Background

Background Senescence-specific cysteine protease (SAG12) is a cysteine protease influenced by cytokinin, auxin and sugars and involved in developmental senescence specific cell death.