Anti-Rad53 antibody (ab104232)
Key features and details
- Rabbit polyclonal to Rad53
- Suitable for: WB
- Reacts with: Saccharomyces cerevisiae
- Isotype: IgG
Overview
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Product name
Anti-Rad53 antibody
See all Rad53 primary antibodies -
Description
Rabbit polyclonal to Rad53 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Saccharomyces cerevisiae
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Immunogen
Synthetic peptide corresponding to Saccharomyces cerevisiae Rad53 aa 800 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab134635)
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab104232 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species WB Saccharomyces cerevisiaeApplication Abreviews Notes WB (3) 1/2000. Detects a band of approximately 92 kDa (predicted molecular weight: 92 kDa). Abcam recommends using milk as the blocking agent (4%)Notes WB
1/2000. Detects a band of approximately 92 kDa (predicted molecular weight: 92 kDa). Abcam recommends using milk as the blocking agent (4%)Target
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Function
Controls S-phase checkpoint as well as G1 and G2 DNA damage checkpoints. Phosphorylates proteins on serine, threonine, and tyrosine. Prevents entry into anaphase and mitotic exit after DNA damage via regulation of the Polo kinase CDC5. Seems to be involved in the phosphorylation of RPH1. -
Sequence similarities
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHEK2 subfamily.
Contains 2 FHA domains.
Contains 1 protein kinase domain. -
Domain
FHA domains are phosphothreonine recognition modules, FHA 1 strongly selects for Asp at position +3 relative to phosphothreonine, whereas FHA 2 selects for Ile in this position. -
Post-translational
modificationsAutophosphorylated. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 855950 Saccharomyces cerevisiae
- SwissProt: P22216 Saccharomyces cerevisiae
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Alternative names
- CHEK2 homolog antibody
- CHK2 homolog antibody
- MEC2 antibody
see all
Images
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Western blot - Anti-Rad53 antibody (ab104232)This image is courtesy of an abreview submitted by Michela Clerici, University of Milano-BiococcaAll lanes : Anti-Rad53 antibody (ab104232) at 1/2000 dilution
Lane 1 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells- TCA prep.
Lane 2 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells treated with 20mg/ml phleomycin for 1 hour - TCA prep
Lane 3 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells- TCA prep
Lane 4 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells arrested in G2 phase with nocodazole- TCA prep.
Lane 5 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells arrested in G2 phase with nocodazole and treated with 20mg/ml phleomycin- TCA prep.
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Amersham anti-rabbit IgG conjugated to HRP at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 92 kDa
Additional bands at: >92 kDa (possible post-translational modification)
Exposure time: 2 minutesBlocking: 4% milk for 1 hour
Phosphorylation of Rad53 induced by phleomycin treatment see PubmedID:152856
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Western blot - Anti-Rad53 antibody (ab104232)This image is courtesy of Ilaria Guerini, Gurdon Institute, United KingdomAll lanes : Anti-Rad53 antibody (ab104232) at 1/2000 dilution
Lane 1 : TCA preps from exponentially growing WT yeast cells.
Lane 2 : TCA preps from exponentially growing WT yeast cells treated with Hydroxyurea (HU).
Lane 3 : TCA preps from exponentially growing rad53delta yeast cells.
Secondary
All lanes : 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 92 kDa
Exposure time: 30 seconds -
Western blot - Anti-Rad53 antibody (ab104232)All lanes : Anti-Rad53 antibody (ab104232) at 1/1000 dilution
Lanes 1 & 5 : Alpha-factor arrested WT cells.
Lane 2 : WT cells 20' after release in S-phase.
Lane 3 : WT cells 40' after release in S-phase.
Lane 4 : WT cells 60' after release in S-phase.
Lane 6 : WT cells 20' after release in S-phase in 200mM HU.
Lane 7 : WT cells 40' after release in S-phase in 200mM HU.
Lane 8 : WT cells 60' after release in S-phase in 200mM HU.
Lanes 9 & 13 : Alpha-factor arrested rad53delta cells.
Lane 10 : rad53delta cells 20' after release in S-phase.
Lane 11 : rad53delta cells 40' after release in S-phase.
Lane 12 : rad53delta cells 60' after release in S-phase.
Lane 14 : rad53delta cells 20' after release in S-phase in 200mM HU.
Lane 15 : rad53delta cells 40' after release in S-phase in 200mM HU.
Lane 16 : rad53delta cells 60' after release in S-phase in 200mM HU.
Secondary
All lanes : Donkey Anti-Rabbit IgG H&L (HRP) (ab16284) at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 92 kDa
Additional bands at: above 92 kDa (possible post-translational modification)
Exposure time: 2 minutes
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Western blot - Anti-Rad53 antibody (ab104232)Western blot showing ab104232 detecting Rad53 in both its unphosphorylated and phosphorylated status. ab104232 was used at 1:2000 dilution.
Datasheets and documents
References (35)
ab104232 has been referenced in 35 publications.
- Liu HW et al. Division of Labor between PCNA Loaders in DNA Replication and Sister Chromatid Cohesion Establishment. Mol Cell 78:725-738.e4 (2020). PubMed: 32277910
- Porcella SY et al. Separable, Ctf4-mediated recruitment of DNA Polymerase a for initiation of DNA synthesis at replication origins and lagging-strand priming during replication elongation. PLoS Genet 16:e1008755 (2020). PubMed: 32379761
- Meng X et al. DNA polymerase e relies on a unique domain for efficient replisome assembly and strand synthesis. Nat Commun 11:2437 (2020). PubMed: 32415104
- Saatchi F & Kirchmaier AL Tolerance of DNA Replication Stress Is Promoted by Fumarate Through Modulation of Histone Demethylation and Enhancement of Replicative Intermediate Processing in Saccharomyces cerevisiae. Genetics 212:631-654 (2019). PubMed: 31123043
- Diernfellner ACR et al. A pathway linking translation stress to checkpoint kinase 2 signaling in Neurospora crassa. Proc Natl Acad Sci U S A 116:17271-17279 (2019). PubMed: 31413202
Images
-
Western blot - Anti-Rad53 antibody (ab104232) This image is courtesy of an abreview submitted by Michela Clerici, University of Milano-BiococcaAll lanes : Anti-Rad53 antibody (ab104232) at 1/2000 dilution
Lane 1 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells- TCA prep.
Lane 2 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells treated with 20mg/ml phleomycin for 1 hour - TCA prep
Lane 3 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells- TCA prep
Lane 4 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells arrested in G2 phase with nocodazole- TCA prep.
Lane 5 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells arrested in G2 phase with nocodazole and treated with 20mg/ml phleomycin- TCA prep.
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Amersham anti-rabbit IgG conjugated to HRP at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 92 kDa
Additional bands at: >92 kDa (possible post-translational modification)
Exposure time: 2 minutesBlocking: 4% milk for 1 hour
Phosphorylation of Rad53 induced by phleomycin treatment see PubmedID:152856
-
Western blot - Anti-Rad53 antibody (ab104232) This image is courtesy of Ilaria Guerini, Gurdon Institute, United KingdomAll lanes : Anti-Rad53 antibody (ab104232) at 1/2000 dilution
Lane 1 : TCA preps from exponentially growing WT yeast cells.
Lane 2 : TCA preps from exponentially growing WT yeast cells treated with Hydroxyurea (HU).
Lane 3 : TCA preps from exponentially growing rad53delta yeast cells.
Secondary
All lanes : 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 92 kDa
Exposure time: 30 seconds
-
Western blot - Anti-Rad53 antibody (ab104232)All lanes : Anti-Rad53 antibody (ab104232) at 1/1000 dilution
Lanes 1 & 5 : Alpha-factor arrested WT cells.
Lane 2 : WT cells 20' after release in S-phase.
Lane 3 : WT cells 40' after release in S-phase.
Lane 4 : WT cells 60' after release in S-phase.
Lane 6 : WT cells 20' after release in S-phase in 200mM HU.
Lane 7 : WT cells 40' after release in S-phase in 200mM HU.
Lane 8 : WT cells 60' after release in S-phase in 200mM HU.
Lanes 9 & 13 : Alpha-factor arrested rad53delta cells.
Lane 10 : rad53delta cells 20' after release in S-phase.
Lane 11 : rad53delta cells 40' after release in S-phase.
Lane 12 : rad53delta cells 60' after release in S-phase.
Lane 14 : rad53delta cells 20' after release in S-phase in 200mM HU.
Lane 15 : rad53delta cells 40' after release in S-phase in 200mM HU.
Lane 16 : rad53delta cells 60' after release in S-phase in 200mM HU.
Secondary
All lanes : Donkey Anti-Rabbit IgG H&L (HRP) (ab16284) at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 92 kDa
Additional bands at: above 92 kDa (possible post-translational modification)
Exposure time: 2 minutes
-
Western blot - Anti-Rad53 antibody (ab104232)Western blot showing ab104232 detecting Rad53 in both its unphosphorylated and phosphorylated status. ab104232 was used at 1:2000 dilution.
