RPB1 | RNA polymerase II subunit B1

AS11 1804 Clonality: Polyclonal Host: Rabbit | Reactivity:Arabidopsis thaliana

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product information
Background		DNA-directed RNA polymerase II subunit 1 (EC=2.7.7.6) catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. This enzyme is the central component of the basal RNA polymerase II transcription machinery. Synonymes:DNA-directed RNA polymerase III largest subunit,DNA-directed RNA polymerase II subunit RPB1.
Immunogen		KLH-conjugated synthetic peptide derived from known higher plant polymerase II sequences, including Arabidopsis thaliana P18616, At4g35800
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Affinity purified serum 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		Immunoprecipitation (IP)
Related products		AS10 710 \| anti-H3 \| histone 3, rabbit antibody collection of antibodies to DN/RNA/cell cycle Plant and algal protein extraction buffer Secondary antibodies
Additional information		Purified IgG was lyophilized from PBS pH 7.4 + 0.02 % sodium azide. Therefore only distilled water is required for its re-constitution.

application information
Recommended dilution		1,2 or 5 µg (IP)
Expected \| apparent MW		204 kDa
Confirmed reactivity		Arabidopsis thaliana
Predicted reactivity		Amygdalus persica, Asparagus officinalis, Brassica rapa, Capsella rubella, Glycine max, Hordeum vulgare, Solanum lycopersicum, Oryza glaberrima, Panicum italicum, Persea americana, Pisum sativum, Ricinus communis, Solanum tuberosum, Sorghum vulgare, Vitis vinifera
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
Additional information
Selected references		Dolata et al. (2015). NTR1 is required for transcription elongation checkpoints at alternative exons in Arabidopsis. EMBO J. 2015 Jan 7. pii: e201489478.

application example

Chromatin immunoprecipitation (ChIP). Chromatin immunoprecipitation was performed as described (Bowler et al. (2004), Plant Journal) with modifications. Sonic and IP buffer were prepared as described (Kaufmann et al. (2010)), Nature Protocols). Chromatin pellet was sonicated at 4 °C with a Diagenode Bioruptor set at high intensity for 10 min (30 sec on, 30 sec off intervals) to obtain 250-500 bp DNA fragments. Chromatin (20 µg) was immunoprecipitated with a RNA Pol II antibody (2µg/IP)and Dynabeads®Protein G (Life Technologies). Reverse corsslinking and elution of DNA fragments were performed using Chelex (Biorad) at 95°C, 10 min as described (Rowley et al. (2013), Methods). Sample without antibody was used to determine the nonspecific background pulled-down directly by the beads. Relative abundance of regions of interest in immunoprecipitated DNA was measured by real-time PCR. Red lines indicates amplified region. Values on the charts are shown as the mean ± SD level (% of input) of RNA Pol II on GAPDH gene (A) and MIR168a (B) gene. Data comes from three independent experiments.

Courtesy of MSc Jakub Dolata, Adam Mickiewicz University, Poland

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