Plant/Algal cell antibodies / Compartment markers / Nuclei

product information

RNA polymerase I is a nuclear located DNA-dependent enzyme involved in RNA elongation and regulation of transcription. In yeast subunit A12.2 has been described as homologous to the Rpb9 subunit from polymerase II.

KLH-conjugated peptide derived from the Arabidopsis thaliana A12.2 (At3g25940) protein sequence. This sequence is only weakly conserved in other eukaryotic sequences available in the databases.

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.

western blot (WB)

to be added when available

This product has previously been labelled as anti-At3g25940 transcription factor S-II (TFIIS) domain-containing protein.

Protocol for isolation of cytosolic and nuclear fractions can be found here.

application information

1 : 2 000 with standard ECL (WB)

13.6

Arabidopsis thaliana

Arabidopsis thaliana

no confirmed exceptions from predicted reactivity known in the moment

This antibody is specific for A12.2 subunit of RNA polymerase I but NOT RNA polymerase II or IV from Arabidopsis thaliana

to be added when available

application example

10 µg of total protein from (1) Arabidopsis thaliana and (4) Oryza sativa leafs extracted with PEB (AS08 300), as well as (2) cytosolic and (3) nuclear fractions of Arbaidopsis thaliana leafs were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-A12.2 (AS07 255, 1:1000, 1h) and secondary anti-rabbit (1:20000, 1 h) antibody (HRP conjugated, Abcam) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with standard ECL (Invitrogen) using a Fuji LAS-3000 CCD (300s, standard sensitivity). The target A12.2 is specifically detected only in the nuclear extract of Arabidopsis thaliana (3).