- Datasheet
- Specific References (2)
- Protocols
Overview
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Product nameAnti-Rad9 antibody
See all Rad9 primary antibodies -
DescriptionRabbit polyclonal to Rad9
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Host speciesRabbit
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Tested applicationsSuitable for: ChIP/Chip, IHC-P, WB, IPmore details
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Species reactivityReacts with: Human
Predicted to work with: Cow, Dog, Pig, Ferret, Rhesus monkey, Gorilla, Bat
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Immunogen
Synthetic peptide corresponding to a region between residue 350 and the C-terminus (residue 391) of Human Rad9 (NP_004575.1)
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Positive control
- Whole cell lysate from HeLa and 293T cells.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.09% Sodium azide
Constituents: 1.815% Tris, 1.764% Sodium citrate, 0.021% PBS -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab70810 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 |
|---|---|---|
| ChIP/Chip | Use at an assay dependent concentration. | |
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| WB | 1/2000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 43 kDa). | |
| IP | Use at 2-5 µg/mg of lysate. |
Target
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FunctionComponent of the 9-1-1 cell-cycle checkpoint response complex that plays a major role in DNA repair. The 9-1-1 complex is recruited to DNA lesion upon damage by the RAD17-replication factor C (RFC) clamp loader complex. Acts then as a sliding clamp platform on DNA for several proteins involved in long-patch base excision repair (LP-BER). The 9-1-1 complex stimulates DNA polymerase beta (POLB) activity by increasing its affinity for the 3'-OH end of the primer-template and stabilizes POLB to those sites where LP-BER proceeds; endonuclease FEN1 cleavage activity on substrates with double, nick, or gap flaps of distinct sequences and lengths; and DNA ligase I (LIG1) on long-patch base excision repair substrates. RAD9A possesses 3'->5' double stranded DNA exonuclease activity. Its phosphorylation by PRKCD may be required for the formation of the 9-1-1 complex.
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Sequence similaritiesBelongs to the rad9 family.
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Post-translational
modificationsConstitutively phosphorylated on serine and threonine amino acids in absence of DNA damage. Hyperphosphorylated by PRKCD and ABL1 upon DNA damage. Its phosphorylation by PRKCD may be required for the formation of the 9-1-1 complex. -
Cellular localizationNucleus.
- Information by UniProt
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Database links
- Entrez Gene: 5883 Human
- Omim: 603761 Human
- SwissProt: Q99638 Human
- Unigene: 655354 Human
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Alternative names
- Cell cycle checkpoint control protein antibody
- Cell cycle checkpoint control protein RAD9A antibody
- DNA repair exonuclease rad9 homolog A antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Rad9 with ab70810 at 1/1000 (1µg/ml). Detection: DAB. -
Western blot - Anti-Rad9 antibody (ab70810)All lanes : Anti-Rad9 antibody (ab70810) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg
Lane 2 : HeLa whole cell lysate at 15 µg
Lane 3 : HeLa whole cell lysate at 5 µg
Lane 4 : 293T whole cell lysate at 50 µg
Developed using the ECL technique.
Predicted band size: 43 kDa
Observed band size: 55 kDa (why is the actual band size different from the predicted?)
Exposure time: 3 minutes -
Immunoprecipitation - Anti-Rad9 antibody (ab70810)1mg whole cell lysate from HeLa cells was immunoprecipitated using ab70810 at 3ug/mg of lysate (lane 1) or a control rabbit Ig (lane 2). For the subsequent western blot, 20% of the immunoprecipitate was loaded per lane, and probed with ab70810 at 1ug/ml.
Detection: chemiluminescence with exposure time of 30 seconds. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)IHC image of ab70810 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab70810, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
References
This product has been referenced in:
- Gong Y et al. PHF11 promotes DSB resection, ATR signaling, and HR. Genes Dev 31:46-58 (2017). WB ; Mouse . Read more (PubMed: 28115467) »
- Panigrahi SK et al. Regulation of NEIL1 protein abundance by RAD9 is important for efficient base excision repair. Nucleic Acids Res 43:4531-46 (2015). Read more (PubMed: 25873625) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"