AS12 1863 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
AS12 1863 Anti-ABI5 | Abscisic acid insensitive 5 (peptide antibody)
Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana ABI5 sequence,UniProt: Q9SJN0, TAIR: AT2G36270 Host: Rabbit Clonality: Polyclonal Purity: Immunogen affinity purified serum in PBS pH 7.4. Format: Lyophilized Quantity: 2x50 µg Reconstitution: For reconstitution add 50 µl of sterile water to each tube Storage: Store lyophilized/reconstituted 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: Immunolocalization (IL), Western blot (WB) Recommended dilution: 1: 140 (IL), 1 : 200 (WB) Expected | apparent MW: 47 kDa
Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Brassica napus, Populus trichocarpa
Species of your interest not listed? Contact usNot reactive in: No confirmed exceptions from predicted reactivity are currently known 2, 5 and 10 ug of total protein (run on separate lanes) extracted from the Columbia ecotype (Arabidopsis thaliana) seeds using Acetone extraction was separated using the Bolt® Bis-Tris Plus Gel system on a 4-12% gradient SDS-PAGE gel, blotted using the turbo-blot system (BIO-RAD) to transfer onto a PVDF membrane (7min). SNAP-ID (Millipore) system was used for blocking and antibody labeling. Blocking occured for 30 minutes (no agitation, 0.05 % skim milk in dest. water). Primary antibody labeling was done for 10 minutes at 1:200 dilution. Followed by 3x10 ml washes (PBST). Then blotted with secondary antibody (anti-rabbit IgG HRP conjugated from Agrisera AS09 602) for 10 min (1:1000 dilution) followed by three washes.Blot was developed using ECL as per manufacturer’s instructions. Exposure time was 2 min.
Courtesy of Nay Chi, Pogson Lab, AustraliaAdditional information: MG132 is recommened to be added to extraction buffer as ABI5 is degraded by proteasome as well as homogenization with thiourea and bead beater.
Protocol for protein extraction from seeds can be requested here.
Additional information (application): ABI5 protein is present in very low levels therefore specific material should be used for analysis as well as chemiluminescence detection reagent in extreme low femtogram range, as AgriseraECLSuperBright. ABI5 (abscisic acid insensitive 5) is involved in ABA-regulated gene expression during seed development and subsequent vegetative stage and acts as the major mediator of ABA repression of growth. Binds to the embryo specification element and the ABA-responsive element (ABRE) of the Dc3 gene promoter and to the ABRE of the Em1 and Em6 genes promoters. Alternative names: ABI5, ABA INSENSITIVE 5, GIA1, GROWTH-INSENSITIVITY TO ABA 1, Dc3 promoter-binding factor 1, AtDPBF1, GROWTH-INSENSITIVITY TO ABA 1, bZIP transcription factor 39, AtbZIP39.