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Rnr3 | Ribonucleoside-diphosphate reductase large chain 2

AS09 574 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Saccharomyces cerevisiae

Rnr3 | Ribonucleoside-diphosphate reductase large chain 2 in the group Antibodies, Bacterial/Fungal at Agrisera AB (Antibodies for research) (AS09 574)

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Product name, number (Agrisera, Sweden)

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Saccharomyces cerevisiae Rnr3 protein sequence UniProt: P21672

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum
Format Lyophilized in PBS pH 7.4
Quantity 100 µg
Reconstitution For reconstitution add 100 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 : 500-1 : 1000 (WB)
Expected | apparent MW

97.5 | 98 kDa

Reactivity

Confirmed reactivity Saccharomyces cerevisiae
Predicted reactivity

Saccharomyces cerevisiae

Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples

Application example

western blot using anti-Rnr3 antibodies

10 μl total protein from 9.25 x 107 cells of Saccharomyces cerevisiae extracted with 20% TCA as described below were separated on 10% SDS-PAGE and blotted 1.5h (0.5 A) to a nitrocellulose membrane (Whatman PROTRAN BA 85, 0.45 μm). Blots were blocked with 5% non-fat dry milk in TBST for 1.5h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 200 overnight at 4C° with agitation. The antibody solution was decanted and the blot was washed 3 times for 10 min in TBST at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, (AS09 602) diluted to 1:50 000 for 1h at RT with agitation. The blot was washed as above and developed for 3 min with chemiluminescent detection reagent, according to the manufacturers instructions. Exposure time was 10 min.

Courtesy Dr. Andrei Chabes, Umeå University, Sweden

Additional information

Load per well was approx 3x10^6 cells (of a total extract).

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Background

Background

Saccharomyces cerevisiae Rnr3 is catalyzing the biosynthesis of deoxyribonucelaotides. Alternative names: Ribonucleotide reductase large subunit 2, ribonucleotide reductase DNA damage-inducible regulatory subunit 2, ribonucleotide reductase R1 subunit 2

Product citations

Selected references Schmidt et al. (2019). Inactivation of folylpolyglutamate synthetase Met7 results in genome instability driven by an increased dUTP/dTTP ratio. Nucleic Acids Res. 2019 Oct 24. pii: gkz1006. doi: 10.1093/nar/gkz1006.
Sampaio-Marques et al. (2019). α-Synuclein toxicity in yeast and human cells is caused by cell cycle re-entry and autophagy degradation of ribonucleotide reductase 1. Aging Cell. 2019 Aug;18(4):e12922. doi: 10.1111/acel.12922.
Lafuente-Barquero et al. (2017). The Smc5/6 complex regulates the yeast Mph1 helicase at RNA-DNA hybrid-mediated DNA damage. PLOS Genetics, December 27, 2017, doi.org/10.1371/journal.pgen.1007136
Schmidt et al. (2017). Alterations in cellular metabolism triggered by URA7 or GLN3 inactivation cause imbalanced dNTP pools and increased mutagenesis. Proc Natl Acad Sci U S A. 2017 May 30;114(22):E4442-E4451. doi: 10.1073/pnas.1618714114.
Graf et al. (2017). Telomere Length Determines TERRA and R-Loop Regulation through the Cell Cycle. Cell. 2017 Jun 29;170(1):72-85.e14. doi: 10.1016/j.cell.2017.06.006.
Williams et al. (2017). The role of RNase H2 in processing ribonucleotides incorporated during DNA replication. DNA Repair (Amst). 2017 Mar 6. pii: S1568-7864(16)30431-1. doi: 10.1016/j.dnarep.2017.02.016.

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