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RPB2 | DNA-directed RNA polymerase II subunit 2

AS20 4376   | Clonality: Polyclonal  |  Host: Chicken | Reactivity: Arabidopsis thaliana

RPB2 | DNA-directed RNA polymerase II subunit 2 in the group Antibodies for Plant/Algal  / Global Antibodies at Agrisera AB (Antibodies for research) (AS20 4376)

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Product Information

Immunogen KLH-conjugated peptide derived from Arabidopsis thaliana RPB2 UniProt: P38420-1, TAIR: AT4G21710
Host

Chicken

Clonality Polyclonal
Purity Affinity purified on antigen, in PBS pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 µl, of sterile 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 : 1000 (WB)
Expected | apparent MW 135 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Arachis duranensis, Brachypodium distachyon, Brassica campestris,Brassica oleracea, Brassica rapa, Capsicum annuum,Cucumis melo, Cucumis sativus, Hordeum vulgare, Homo sapiens, Lupinus angustifolius, Malus baccata, Nicotiana tabacum, Oryza sativa, Petunia hybrida, Phaseolus vulgaris, Populus alba, Raphanus sativus, Sorghum bicolor, Trifolium subterraneum, Triticum aestivum, Zea mays
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Western blot using anti-RPB2 antibodies

10-40 µg/well of total protein extracted from 7-d-old Arabidopsis thaliana seedlings (grown in continuous light at 22ºC) with 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.25% Triton X-100, 2 mM PMSF and 1x protease inhibitor cocktail (Roche) and denatured with 1XLaemli buffer at 100°C for 5 min, were separated in duplicate on 7% SDS-PAGEs and blotted 1h 15 min to PVDF (0.2 um), using wet transfer. Blots were blocked with TBS-T/5% milk for 1h/RT with agitation. Blots were incubated in the primary antibodies at a dilution of 1:1000 (anti-NRPB2) ON/4°C in TBS-T/5% milk with agitation. The anti-NRPB2 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 (rabbit anti-chicken IgG horse radish peroxidase conjugated, AS10 1489) diluted to 1:1000 in for 1h/RT with agitation. The two blots were washed as above and developed for 5 min with chemiluminescent detection reagent, following manufacture's recommendations.  Exposure time was 10 seconds (ImageQuant 800, 3x3 bins; Amersham).

Courtesy of Alberto Palacios-Abella from Dr. David Alabadi lab, CSIC-UPV, Spain

Additional information

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Background

Background RPB2 (DNA-directed RNA polymerase II subunit 2) is a unique second-largest subunit of DNA-dependent RNA polymerase II.  Alternative names: DNA-directed RNA polymerase II 135 kDa polypeptide DNA-directed RNA polymerase II subunit RPB2 (EC:2.7.7.6), RNA polymerase II subunit B2, Protein EMBRYO DEFECTIVE 1989

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