AS22 4789 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Amaranthus cruentus, Arabidopsis thaliana, Beta vulgaris
AS22 4789 Anti-AHA2 p | ATPase 2 phosphorylated, plasma membrane-type
Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana AHA2 protein sequence UniProt: P19456, TAIR: At4g30190 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 or deionized 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 : 10 000 (WB) Expected | apparent MW: 104.4 | kDa Confirmed reactivity: Amaranthus cruentus, Arabidopsis thaliana, Beta vulgaris Predicted reactivity: Brassica oleracea, Camelina sativa, Capsella rubella, Eutrema salsugineum, Raphanus sativus
Species of your interest not listed? Contact usNot reactive in: No confirmed exceptions from predicted reactivity are currently known Samples:
1 - Recombinant protein MBP-C terminal region of AHA2, phosphorylation treatment
2 - Recombinant protein MBP-C terminal region of AHA2, subsequently dephosphorylated
3 - Recombinant protein MBP, phosphorylation treatment
4 - Recombinant protein MBP-C terminal region of AHA2, Thr947Ala, phosphorylation treatment 5- Mark: MW markers1 µg/well of recombinant protein produced in E. coli and purified by affinity chromatography was incubated in the reaction mixture (30 ul). 3 ul of the reaction were mixed with 6 ul Laemli 2X and denatured at 95°C for 5 min. Samples were separated on 10% SDS-PAGE and blotted overnight (ON) to PVDF (Inmobilon®-FL) (pore size of 0.45 µm) using wet transfer. The blot was blocked with 3 % non-fat milk 2h/RT with agitation. The blot was incubated in the primary antibody at a dilution of 1:10 000 for 90 min/RT with agitation 3% non-fat milk TBS 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. The blot was incubated in a matching secondary antibody (anti-rabbit IgG horseradish peroxidase conjugated) diluted to 1: 5000 for 90 min/RT with agitation. The blot was washed as above and developed with the following chemiluminescent detection reagent: AgriseraBright. Exposure time was 2 min.
Courtesy of Cristian Mayordomo, IBMCP, Universidad Politécnica de Valencia, Spain
Titration of the AS22 4789: AHA2 p | ATPase 2 phosphorylated, plasma membrane-type antibody. Ten µg of protein from Amaranth stem mixed membranes (TO), or from beetroot plasma membranes either pre-phosphorylated, i.e. treated with ATP (MP+P), or not (MP* and MP-P) were blotted and probed with the indicated dilutions of antibody. Molecular mass markers (MPM) have, from top to bottom 250, 130, 100, 70 (red) and 55 kDa, respectively.
Courtesy Dr. Luis González, CINVESTAV, Mexico
AGO expression may be cell/tissue specific and using floral tissue is recommended where most of the AGOs are expressed the highest. Seedlings can be used as a negative control.
Use of proteasome inhibitors as MG132 can help to stabilize AGO proteins during extraction procedure.
A recommended whole-mount immunolocalization protocol can be found here.The plasma membrane H+ ATPase of plants and fungi generates a proton gradient that drives the active transport of nutrients by H+-symport