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
Suitable for: WBmore details -
Species reactivity
Reacts with: Saccharomyces cerevisiae -
Immunogen
Synthetic peptide corresponding to Saccharomyces cerevisiae Rad53 aa 800 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab134635) -
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research 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.
Application | Abreviews | Notes |
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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%)
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Notes |
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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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All 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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All 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 -
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: 92 kDa (possible post-translational modification)
Exposure time: 2 minutes
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Western blot showing ab104232 detecting Rad53 in both its unphosphorylated and phosphorylated status. ab104232 was used at 1:2000 dilution.
Datasheets and documents
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SDS download
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Datasheet download
References (59)
ab104232 has been referenced in 59 publications.
- Masłowska KH et al. Eukaryotic stress-induced mutagenesis is limited by a local control of translesion synthesis. Nucleic Acids Res 50:2074-2080 (2022). PubMed: 35104879
- Conti MM et al. Repression of essential cell cycle genes increases cellular fitness. PLoS Genet 18:e1010349 (2022). PubMed: 36037231
- Reusswig KU et al. Unscheduled DNA replication in G1 causes genome instability and damage signatures indicative of replication collisions. Nat Commun 13:7014 (2022). PubMed: 36400763
- Sheu YJ et al. Prevalent and dynamic binding of the cell cycle checkpoint kinase Rad53 to gene promoters. Elife 11:N/A (2022). PubMed: 36520028
- Casari E et al. Dpb4 promotes resection of DNA double-strand breaks and checkpoint activation by acting in two different protein complexes. Nat Commun 12:4750 (2021). PubMed: 34362907