AS18 4235 | Clonality: Polyclonal | Host: Rabbit | Reactivity: higher plants
AS18 4235 Anti-AHB1 | Non-symbiotic hemoglobin 1 (other species)
Immunogen: KLH-conjugated peptide derived from protein sequences of AHB1 from: Ricinus communis UniProt: B9RZD8, Solanum lycopersicum UniProt: Q9AWA9, Solanum tuberosum UniProt: Q8GV42 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. Re-constituted antibody can be stored at 4°C for several days to weeks.
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: Medicago sativa Predicted reactivity: Cannabis sativa, Capsicum annuum, Gossypium sp., Manihot esculenta, Nicotiana tabacum, Physcomitrium patens, Solanum lycopersicum, Solanum tuberosum, Ricinus communis, Vitis vinifera Not reactive in: No confirmed exceptions from predicted reactivity are currently known 20 or 10 µg/well of total protein extracted freshly from roots from 5 days seedlings (flooded for or without flooding – control) with write exact buffer components 0.1 M Tris-HCl 7,5, 10 % glycerol, 1 mM EDTA, 2 mM DTT, 1 mM PMSF, 2 % PVPP and denatured with 62,5 mM Tris-HCl pH 6.8, 2 % SDS, 10 % glycerol, 20 mM DTT, 0,01 % bromophenol blue, at 98°C or 70°C for 5 min. Proteins were separated on 10 % SDS-PAGE and blotted 2h (constant 90 V, 4°C) to PVDF/nitrocellulose (pore size of 0,2 um), using wet, semi-dry or dry transfer. Blot was blocked with 5 % milk or % BSA for 1h/RT or 4°C/ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h/RT with agitation in TBS-T or ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in for 1h/RT with agitation. The blot was washed as above and developed for min with Agrisera ECLBright or AgriseraECLSuperBright with BioRad ChemiDoc XR S+. Exposure time was 40 seconds.
Courtesy of Dr. Paweł Staszek, SGGW, Warsaw, Poland
For detection in plant samples, plants need to be flooded as AHB1 levels in non-treated plants are very low.
The antibody is binding to human hemoglobin.AHB1 (Non-sybmiotic hemoglobin) plays an important role in as nitric oxide scavenger and stress response mediator.