AS18 4230 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
AS18 4230 Anti-AHB1 | Non-symbiotic hemoglobin 1 (A. thaliana)
Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana AHB1, UniProt: O24520, TAIR: AT2G16060 Host: Rabbit Clonality: Polyclonal Purity: Antigen affinity purified serum, in PBS pH 7.4. Format: Lyophilized Quantity: 50 µg Reconstitution: For reconstitution add 50 µl, of sterile water. Storage: Lyophilized antibody can be stored at -20°C for up to 3 years. 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 : 1000 (WB) Expected | apparent MW: kDa Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Malus domestica Not reactive in: No confirmed exceptions from predicted reactivity are currently known Samples:
1 - 90 ug of Arabidopsis thaliana whole seedling extract, Col-0 (wild type) submerged to induce PGB1
2 - 90 ug of Arabidopsis thaliana whole seedling extract, Col-3 (wild type) submerged to induce PGB1
3 - 90 ug of Arabidopsis thaliana whole seedling extract, Col-0; 35S: GFP-PGB1 in air (control)
4 - 90 ug of Arabidopsis thaliana whole seedling extract, C24; 35S:PGB1 in air (control)
5 - 90 ug of Arabidopsis thaliana whole seedling extract, C24; 35S:PGB1 submerged to induce PGB1
6 - 90 ug of Arabidopsis thaliana whole seedling extract, Col-0 (wild type in air (control)
7 - 90 ug of Arabidopsis thaliana whole seedling extract, C24 (wild type) in air (control)
8 - 90 ug of Arabidopsis thaliana whole seedling extract, knock down mutant of PGB1 (C24 background) submerged to induce PGB1
9 - 90 ug of Arabidopsis thaliana whole seedling extract, knock out mutant of PGB1 (Col-3 background) submerged to induce PGB1 (AHB1)
Mark: MW markers: Dual Color (BioRad)90 µg/well of total protein extracted freshly from Arabidopsis thaliana seedlings with extraction buffer (100mM Tris-HCl, 150mM NaCl, 0.25% NP-40) containing 1mM PMSF and 1X cOmplete® EDTA-free proteases inhibitors, Sigma) and denatured with buffer (Tris-HCl 0,25 M, pH 6,8; SDS 8% (w/v); glicerol 40% (v/v); β-Mercaptoethanol 20% (v/v) and Bromophenol Blue) at 90°C for 10 min were separated on 12% SDS-PAGE and blotted (20min; 2A; 25V) to PVDF (pore size of 0.2 µm), using semi-dry transfer. Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 ON/4°C with agitation. The antibody solution was decanted and the blot was washed 2 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (Goat anti-rabbit IgG HRP peroxidase conjugated, AS09 602) diluted to 1:25 000 in for 1h/RT with agitation. The blot was washed 4 times for 5 min in TBS-T at RT with agitation and developed with Agrisera ECL SuperBright, AS16 ECL-S. Exposure time was 5 seconds.
The lines used in the experiments were generated by:
-Hebesltrup et al., 2008 corresponds to 35S: GFP-PGB1
-Perazzolli et al., 2004 corresponds to 35S:PGB1
Courtesy of Dr. Inmaculada Sánchez-Vicente and Isabel Manrique-Gil, USAL-CIALE Salamanca, SpainConditions for detection of endogenous AHB1 remain to be determined.
AHB1 (Non-symbiotic hemoglobin 1) is involved in a cellular response to hypoxia and displays oxygen carrier activity. AHB1 is induced during flooding in roots. It is also induced in seeds during germination and in meristematic tissues bas they experience oxygen depletion.
Alternative names: ARAth, GLB1, Hb1AHB1, ARATH GLB1, ATGLB1, CLASS I HEMOGLOBIN, GLB1, HB1, HEMOGLOBIN 1, NSHB1, PGB1, PHYTOGLOBIN 1.