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Invitrogen

RAD50 Polyclonal Antibody

5个已发表图片

4 篇参考文献

查看其他19个RAD50抗体

货号 PA5-32176

CNY 4,905.00

100 µL

批量或定制请求

抗体检测数据 (1)
已发表图片 (5)

Application

RAD50 Antibody in Western Blot (WB)

图: 1 / 6

RAD50 Antibody (PA5-32176) in WB

Western Blot using RAD50 Polyclonal Antibody (Product # PA5-32176). Sample (10 µg of whole cell lysate). Lane A: Yeast . 5% SDS PAGE. RAD50 Polyclonal Antibody (Product # PA5-32176) diluted at 1:1,000.

RAD50 Antibody in Western Blot (WB)

RAD50 Antibody in Western Blot (WB)

RAD50 Antibody in Western Blot (WB)

RAD50 Antibody in Western Blot (WB)

RAD50 Antibody in Western Blot (WB)

RAD50 Antibody in Western Blot (WB)

Product Image Gallery

RAD50 Antibody in Western Blot (WB) — Western Blot using RAD50 Polyclonal Antibody (Product # PA5-32176). Sample (10 µg of whole cell lysate). Lane A: Yeast . 5% SDS PAGE. RAD50 Polyclonal Antibody (Product # PA5-32176) diluted at 1:1,000.

RAD50 Antibody in Western Blot (WB) — Foci formation and DNA repair require MRX oligomerization in vivo. a Serial dilution of exponentially growing S. cerevisiae rad50 ^ ^ cells expressing Rad50 wt/ho/lo -yEVenus, spotted on agar plates (YPD) +/- DNA damage-inducing camptothecin (CPT) or hydroxyurea (HU), and incubated for 2 days at 30 degC. n = 3. b Doubling times of exponentially growing rad50 ^ ^ cells and rad50 ^ ^ strains expressing Rad50 wt/ho/lo -yEVenus +/- CPT treatment. Mean and minimum to maximum values shown, n = 3 independent experiments. c Anti-Rad50 western blot showing galactose-induced overexpression of untagged Rad50 wt/ho/lo in wild-type cells (+Gal-OE). Center panel is contrast-enhanced to show endogenous Rad50 wt levels (wild-type, -Gal-OE). alpha -Tubulin controls equal loading. n = 3. d Serial dilution of exponentially growing wild-type cells expressing galactose-inducible Rad50 wt/ho/lo , spotted on SD-URA agar plates with glucose (-) or galactose (+ variant overexpression), and incubated at 30 degC for 2 days. Before overexpression, cells carrying Rad50 wt/ho/lo were similarly resistant to DNA damage as wild-type cells due to endogenous Rad50 wt (-, top). n = 3. e Representative live-cell images of untreated or CPT-treated rad50 ^ ^ cells expressing Rad50 wt/ho/lo -yEVenus with histone Hta2-mCherry as nuclear marker. Arrows indicate nuclear Rad50 foci (Supplementary Fig. 7a ). f Anti-Rad50 western blot showing expression levels of Rad50 wt/ho/lo -yEVenus in rad50 ^ ^ cells before (-) and

RAD50 Antibody in Western Blot (WB) — Fig. 4 MR oligomerization influences DNA binding behavior and endonuclease activity. a Quantification of EMSAs of MR wt/ho/lo binding to 100 bp-long DNA as in Supplementary Figs. 2a and 5f . Mean +- SEM, n = 3 independent experiments, sigmoidal fit. b TEM of MR ho/lo binding to plasmid DNA. Images representative of n = 3 experiments. MR lo binds dispersedly on DNA without oligomerization, while MR ho dimers oligomerize on DNA to a denser "worm"-like structure compared to the "pearls-on-a-string" assembly of MR wt (Fig. 1b-ii ) at similar conditions (see "Methods"). c TEM of Mre11 wt and Rad50 wt/ho/lo binding to plasmid DNA. n = 3. In contrast to purified Mre11 wt , Rad50 wt forms "pearls-on-a-string"-like structures on linear plasmid DNA. Rad50 lo fails to oligomerize, thus binds dispersedly on DNA, while Rad50 ho clusters into large oligomers on DNA, similarly to MR ho . d, e Endonuclease assays of MR wt/ho/lo -pSae2 ( d , Supplementary Fig. 6a ) with quantification ( e ). Images representative of n = 3 experiments. Mean +- SEM, unpaired two-tailed t -tests. Red star marks the position of the radio-label. f , g Representative endonuclease activity kinetics of MR wt/ho -pSae2 ( f ) with quantification ( g ). Mean +- SEM, n = 3 independent experiments. Red star marks the position of the radio-label. h Representative resection assay of MR ho with probe (green box in cartoon) binding to the 3'-overhang produced. n = 2. Note that the large MR ho oligomers in Fig. 4b are

RAD50 Antibody in Western Blot (WB) — Fig. 2 The MIN motif of Rif2 counteracts the action of the MRX complex in promoting telomere elongation and promotes binding to Rad50. a Southern blot analysis of telomere length of Saccharomyces cerevisiae strains of the indicated genotypes. Genomic DNA was digested with Xho I and detected using a telomeric probe. 'Delta' indicates null alleles obtained by full deletion of the coding sequence. The rif2-min allele carries an F8E P10N substitution. b Viability assays of strains overexpressing a fusion protein containing the first 34 amino acids of Rif2 under control of the galactose-inducible promoter, either in wild-type form (MIN) or mutant (MINmut, carrying the double F8E P10N substitution). Strains were grown in glucose-containing rich medium (YPAD) before plating onto either glucose (uninduced, top row) or induced (galactose, bottom row) plates containing the indicated chemicals and incubated at 30 degC for 2-3 days. c Protein interaction assays with immobilised wild-type or mutant Rif2 (Rif2-min) as bait, and Rad50, Mre11 or Xrs2 proteins used individually as prey. The bound proteins were separated by electrophoresis and detected by Western blotting (for Rad50, anti-Rad50; for Mre11-his, anti-his; and for Xrs2-FLAG, anti-FLAG), or by Ponceau S staining (for Rif2 wild-type or Rif2-min). CTRL lane is prey protein incubated with amylose resin without bait. A representative blot is shown, as well as a quantification of 2-3 independent ones, as indicated. Source data are prov

RAD50 Antibody in Western Blot (WB) — Fig. 5 Rad50S (Rad50-K81I) fails to interact with the MIN motif of Rif2. a Southern blot analysis of telomere length of Saccharomyces cerevisiae strains of the indicated genotypes, carried out as described in Fig. 2a . A quantification of hybridization signals from this gel obtained with Image Gauge v4.1 (Fuji) is shown on the right panel; strains of identical genotypes were averaged together. b Yeast two-hybrid assays of the S. cerevisiae fusion proteins indicated. Plates are minimal medium lacking the nutrients indicated on top. Five-fold serial dilutions were spotted left to right and incubated for 3-4 days at 30 degC. c Protein interaction assays with immobilized wild-type or mutant Rif2 (Rif2-min) as bait, and Rad50 or Rad50-K81I (Rad50S) proteins as prey. The bound proteins were separated by electrophoresis and detected by Western blotting (with anti-Rad50 antibody) or by Ponceau S staining (for Rif2 wild-type or Rif2-min). A representative blot is shown, as well as a quantification of three independent ones, as indicated. Data are presented as mean with SEM. d Viability assays of strains overexpressing a fusion protein containing the first 34 amino acids of Rif2 under control of the galactose-inducible promoter, either in wild-type form (MIN) or mutant (MINmut), as in Fig. 2b . Uninduced strains were plated onto either glucose (uninduced, top row) or induced (galactose, bottom row) medium containing the indicated chemicals and incubated at 30 degC for 2-3 days. e Viabi

RAD50 Antibody in Western Blot (WB) — Figure 6 The N18S mutation weakens the Rad50-Rif2 interaction and is located close to the rad50-s mutations (A) Recombinant MBP-Rif2 was immobilized on amylose resin (lanes 1 and 2). Subsequently, MR or MR N18S was added, as indicated. The resin with bound proteins was washed and eluted with maltose, and the MBP tag on Rif2 was cleaved with PreScission protease. The proteins in the eluate were analyzed by Ponceau (Rif2) and western blotting (anti-Rad50 and anti-His to detect Mre11-His). (B) Comparison of E. coli sbcC (Mre11 ortholog, pink) and sbcD (Rad50 ortholog, blue) in the cutting state (PDB: 6S85 ) and S. cerevisiae Mre11 and Rad50 models, obtained by homology modeling, which are positioned by structural alignment to mimic the same cutting-state conformation. The Rad50 residues (K6, R20, and K81) involved in Sae2 interaction and affected by the rad50-s mutations are shown as orange spheres. The Mre11 residue (T188) affected by the mre11-s mutation is shown as green spheres. The Rad50 residue N18 is drawn in gray spheres, whereas the ADP nucleotide is in yellow. (C) Protein extracts from exponentially growing xrs2 Delta cells were analyzed by western blotting with anti-HA and anti-Myc antibodies either directly (total) or after immunoprecipitation (IP) with anti-Myc antibody.

产品信息

PA5-32176

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1:500-1:3,000

查看4篇已发表文章 4篇已发表文章

产品规格

种属反应

Yeast

已发表种属

Insect, Yeast

宿主/亚型

Rabbit / IgG

分类

Polyclonal

类型

Antibody

抗原

Recombinant fragment corresponding to a region within amino acids 644 and 989 of Yeast RAD50

查看免疫原

偶联物

Unconjugated

形式

Liquid

浓度

1 mg/mL

纯化类型

Antigen affinity chromatography

保存液

0.1M tris glycine, pH 7, with 20% glycerol

内含物

0.01% thimerosal

保存条件

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

运输条件

Wet ice

RRID

AB_2549649

产品详细信息

Recommended positive controls: Yeast.

Predicted reactivity: Yeast (100%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

靶标信息

Rad50-13B3 recognizes human Rad50, a 153a protein involved in DNA double strand break repair. Rad50 is associated with Mre11 and p95 (Nibrin) to form a multiprotein complex involved in the double strand break repair process. Upon DNA damage, Rad50 and Mre11 redistribute and are co-localized within the nucleus of damaged cells.

⚠WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause birth defects or other reproductive harm. For more information go to www.P65Warnings.ca.gov.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

Product References

Mre11-Rad50 oligomerization promotes DNA double-strand break repair.

Nature communications

Kissling VM,Reginato G,Bianco E,Kasaciunaite K,Tilma J,Cereghetti G,Schindler N,Lee SS,Guérois R,Luke B,Seidel R,Cejka P,Peter M

Published figure using RAD50 polyclonal antibody (Product # PA5-32176) in Western Blot

Mon May 02 00:00:00 UTC 2022

Inhibition of MRN activity by a telomere protein motif.

Nature communications

Khayat F,Cannavo E,Alshmery M,Foster WR,Chahwan C,Maddalena M,Smith C,Oliver AW,Watson AT,Carr AM,Cejka P,Bianchi A

Published figure using RAD50 polyclonal antibody (Product # PA5-32176) in Western Blot

Tue Jun 22 00:00:00 UTC 2021

Inhibition of MRN activity by a telomere protein motif.

Nature communications

Khayat F,Cannavo E,Alshmery M,Foster WR,Chahwan C,Maddalena M,Smith C,Oliver AW,Watson AT,Carr AM,Cejka P,Bianchi A

Published figure using RAD50 polyclonal antibody (Product # PA5-32176) in Western Blot

Tue Jun 22 00:00:00 UTC 2021

Sae2 and Rif2 regulate MRX endonuclease activity at DNA double-strand breaks in opposite manners.

Cell reports

Marsella A,Gobbini E,Cassani C,Tisi R,Cannavo E,Reginato G,Cejka P,Longhese MP

Published figure using RAD50 polyclonal antibody (Product # PA5-32176) in Western Blot

Tue Mar 30 00:00:00 UTC 2021

Sae2 and Rif2 regulate MRX endonuclease activity at DNA double-strand breaks in opposite manners.

Cell reports

Marsella A,Gobbini E,Cassani C,Tisi R,Cannavo E,Reginato G,Cejka P,Longhese MP

Published figure using RAD50 polyclonal antibody (Product # PA5-32176) in Western Blot

Tue Mar 30 00:00:00 UTC 2021

Regulatory control of DNA end resection by Sae2 phosphorylation.

Nature communications

Cannavo E,Johnson D,Andres SN,Kissling VM,Reinert JK,Garcia V,Erie DA,Hess D,Thomä NH,Enchev RI,Peter M,Williams RS,Neale MJ,Cejka P

PA5-32176 was used in Western Blotting to define how phosphorylation controls the initiation of DNA end resection and therefore the choice between the key DNA double-strand break repair mechanisms.

Mon Oct 01 00:00:00 UTC 2018

生物信息学

蛋白别名: 153 kDa protein; DNA repair protein RAD50; hsRAD50; RAD50 2; RAD50 PEN; RAD50-2; RAD50-3

基因别名: N0872; RAD50; YNL250W

UniProt ID: (Baker's yeast) P12753

Entrez Gene ID: (Baker's yeast) 855471

nucleotide binding double-stranded DNA binding double-stranded telomeric DNA binding adenylate kinase activity ATP binding hydrolase activity ATPase activity telomeric DNA binding single-stranded telomeric DNA binding metal ion binding G-quadruplex DNA binding ATP-dependent DNA damage sensor activity

参与通路

telomere maintenance via recombination telomere maintenance DNA repair base-excision repair double-strand break repair double-strand break repair via nonhomologous end joining cellular response to DNA damage stimulus telomere maintenance via telomerase synapsis negative regulation of endodeoxyribonuclease activity DNA duplex unwinding maintenance of DNA trinucleotide repeats nucleoside monophosphate phosphorylation meiotic cell cycle chromosome organization involved in meiotic cell cycle nucleic acid phosphodiester bond hydrolysis mitochondrial double-strand break repair via homologous recombination

1x1 image pixel for data collection